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Effects of Site-Mutagenesis of an Amino Acid Triplet Repeat at $M_1$ and $M_2$ Muscarinic Receptors on Receptor Function ($M_1$$M_2$ 무스카린성 수용체에서 아미노산 Triplet Repeat의 Site-Mutagenesis가 수용체기능에 미치는 영향)

  • Lee, Seok-Yong;Lee, Sang-Bok
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.311-321
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    • 1996
  • Both $M_1$ and $M_2$ muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T) at C-terminus ends of the second putative transmembrane domains. This triplet is repeated as LYT-LYT in $M_2$ receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposed fashion (LYT-TYL) in the sequence of $M_1$ receptors. In this work, we employed site-directed mutagenesis to investigate the possible significance of this unique sequence diversity for determining the distinct differential cellular function at the two receptor subtypes. Mutation of the LYTTYL sequence of $M_1$ receptors to the corresponding $M_2$ receptor LYTLYT sequence did not result in a significant change in the binding affinity of the agonist carbachol. The reverse mutation at the $M_2$ receptor also did not modify agonist affinity. Surprisingly, the LYTLYT $M_1$ receptor mutant demonstrated markedly enhanced coupling to activation of phospholipase C without a change in its coupling to increased cyclic AMP formation. There was also an enhanced receptor sensitivity in transducing elevation of intracellular $Ca^{2+}$. On the other hand, the reverse $LYTLYT{\rightarrow}LYTTYL$ mutation in the $M_2$ receptor did not alter its coupling to inhibition of adenylate cyclase, but slightly enhanced its coupling to stimulation of phosphoinositide (PI) hydrolysis. Our data suggest that the LYTTYL/LYTLYT sequence differences between $M_1$ and $M_2$ muscarinic receptors are not important for specifying ligand binding and coupling of various subtypes of muscarinic receptors to different cellular signaling pathways although they might play a role in the modulation of muscarinic reseptor coupling to PI hydrolysis.

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A study on the CRM strategy for medium and small industry of distribution (중소유통업체의 CRM 도입방안에 관한 연구)

  • Kim, Gi-Pyoung
    • Journal of Distribution Science
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    • v.8 no.3
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    • pp.37-47
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    • 2010
  • CRM refers to the operating activities that always maintain and promote good relationship with customers to ultimately maximize the company's profits by understanding the value of customers to meet their demands, establishing a strategy which may maximize the Life Time Value and successfully operating the business by integrating the customer management processes. In our country, many big businesses are introducing CRM initiatively to use it in marketing strategy however, most medium and small sized companies do not understand CRM clearly or they feel difficult to introduce it due to huge investment needed. This study is intended to present CRM promotion strategy and activities plan fit for the medium and small sized companies by analyzing the success factors of the leading companies those have already executed CRM by surveying the precedents to make the distributors out of the industries have close relation with consumers to overcome their weakness in scale and strengthen their competitiveness in such a rapidly changing and fiercely competing market. There are 5 stages to build CRM such as the recognition of the needs of CRM establishment, the establishment of CRM integrated database, the establishment of customer analysis and marketing strategy through data mining, the practical use of customer analysis through data mining and the implementation of response analysis and close loop process. Through the case study of leading companies, CRM is needed in types of businesses where the companies constantly contact their customers. To meet their needs, they assertively analyze their customer information. Through this, they develop their own CRM programs personalized for their customers to provide high quality service products. For customers helping them make profits, the VIP marketing strategy is conducted to keep the customers from breaking their relationships with the companies. Through continuous management, CRM should be executed. In other words, through customer segmentation, the profitability for the customers should be maximized. The maximization of the profitability for the customers is the key to CRM. These are the success factors of the CRM of the distributors in Korea. Firstly, the top management's will power for CS management is needed. Secondly, the culture across the company should be made to respect the customers. Thirdly, specialized customer management and CRM workers should be trained. Fourthly, CRM behaviors should be developed for the whole staff members. Fifthly, CRM should be carried out through systematic cooperation between related departments. To make use of the case study for CRM, the company should understand the customer and establish customer management programs to set the optimal CRM strategy and continuously pursue it according to a long-term plan. For this, according to collected information and customer data, customers should be segmented and the responsive customer system should be designed according to the differentiated strategy according to the class of the customers. In terms of the future CRM, integrated CRM is essential where the customer information gathers together in one place. As the degree of customers' expectation increases a lot, the effective way to meet the customers' expectation should be pursued. As the IT technology improved rapidly, RFID (Radio Frequency Identification) appears. On a real-time basis, information about products and customers is obtained massively in a very short time. A strategy for successful CRM promotion should be improving the organizations in charge of contacting customers, re-planning the customer management processes and establishing the integrated system with the marketing strategy to keep good relation with the customers according to a long-term plan and a proper method suitable to the market conditions and run a company-wide program. In addition, a CRM program should be continuously improved and complemented to meet the company's characteristics. Especially, a strategy for successful CRM for the medium and small sized distributors should be as follows. First, they should change their existing recognition in CRM and keep in-depth care for the customers. Second, they should benchmark the techniques of CRM from the leading companies and find out success points to use. Third, they should seek some methods best suited for their particular conditions by achieving the ideas combining their own strong points with marketing. Fourth, a CRM model should be developed that will promote relationship with individual customers just like the precedents of small sized businesses in Switzerland through small but noticeable events.

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Transection of the posterior horn of the medial meniscus at the posterior tibial attachment - Clinical features and A new repair technique (Pullout suture) - (내측 반월상 연골 후각의 후방 경골 부착부위의 절단 파열 - 임상 양상 및 새로운 봉합 수기(pullout suture) -)

  • Ahn, Jin-Hwan;Ha, Chul-Won;Kim, Ho;Kim, Sung-Min
    • Journal of the Korean Arthroscopy Society
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    • v.3 no.2
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    • pp.109-114
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    • 1999
  • Purpose : The importance of meniscal repair is well recognized. But transeciton of the posterior horn of the medial meniscus at the posterior tibial attachment is rarely documented and known irreparable. We experienced 9 cases of transection, and present clinical features and pull out suture technique. Methods and Materials : There were 9cases of transection of the posterior horn of medial meniscus from September 1998 to July 1999 in our hospital. Age was 59.3 years in average and ranged from 38 to 70years. Clinical features and MRI made diagnosis in all cases. We confirmed the diagnosis with arthroscopy and repaired the transection with pullout suture technique. Clinical features : Transection of the posterior horn of the medial meniscus at the posterior tibia attachment occurred frequently in middle aged people. They complained posterior knee pain, but they have no history of definitive trauma. Characteristically they had difficulty in full flexion of the knee and in having a squatting position. MRI is very important in diagnosis of transection, especially in coronal view, there is separation of the posterior horn of the meniscus from the posterior tibial attachment. Surgical technique : Pullout suture technique includes debridement of fibrous or scar tissue, exposure of the subchondral bone of the posterior tibial attachment site, suture the transected end of the meniscus with PDS suture, bone tunnel formation from the anteromedial aspect of the proximal tibia, insertion of wire loop through the tibia tunnel, pull the PDS suture through the tibia tunnel out of the joint and stabilize the PDS with post-tie technique to the proximal tibia. Conclusion : Transection of the posterior horn of the medial meniscus at the posterior tibial attachment is not common clinically and rarely documented. Clinical features and MRI are very important in diagnosis of this type tear. Arthroscopic pullout sutures is useful for treatment of this type tear of the meniscus.

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A Study on $Na^+$ and Water Reabsorption in the Nephron Segment Beyond Proximal Tubule Measured by Lithium Clearance

  • Han, O-Soo;Goo, Yong-Sook;Sung, Ho-Kyung
    • The Korean Journal of Physiology
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    • v.25 no.2
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    • pp.189-200
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    • 1991
  • During the past few years it has been proposed that lithium clearance can be used as a reliable measure for the outflow of tubular fluid from the proximal tubule. This study was aimed to characterize the inflow dependent reabsorption of Na in renal tubule beyond the proximal tubule. For this purpose, lithium clearance was used as a measure for the inflow from the proximal tubule and the changes in reabsorption fraction of Na and water were determined in rabbits. Rabbits were pretreated with hypotonic saline solutions for an hour (50 mM/L NaCl, 20 ml/hr/kg). And then a hypertonic solution of 500 mM/L NaCl (20 ml/kg) was administered intraperitoneally in conjunction with a bolus of LiCl solution (2 mM/kg, i.v.) for conditioning the $C_{Li}$ and urine flow rate. To rule out the effect of $Li^+$ on tubular functions, a bolus of NaCl solution (2 mM/kg, i.v.) was administered. Fifteen, thirty, and sixty minutes after injection of hypertonic saline arterial blood and urine samples were taken. Urinary and plasma concentrations as well as urinary output of $Li^+,\;Na^+\;and\;K^+$ were measured. From these $C_{Li},\;C_{Na}$ and the reabsorption fraction of Na and water $(Fr_{Na}\;&\;FrH_2O)$ were calculated. These results were compared with those from control groups in which the same amount of isotonic saline (145 mM/L NaCl) and of 15% dextran solution were administered in the same way as that in experimental group. Followings are the results obtained. 1) The plasma concentration of $Na^+$ in rabbits injected with hypertonic saline reached the peak value after 15 min and thereafter no significant change was observed. Hematocrit values did not show any change, while urinary excretion of $Na^+$ increased markedly during the first 15 min and decreased thereafter. These results were not affected by an injection of a small amount of LiCl. 2) The clearances of $Li^+,\;Na^+\;and\;K^+$ in rabbits injected with hypertonic saline and LiCl solution decreased. 3) In spite of the variation in $C_{Li},\;Fr_{Na}$ did not show any significant change while $FrH_2O$ increased gradually. 4) $C_{Li}$ decreased also in rabbits received isotonic saline. $Fr_{Na}$ tended to be higher than that in hypertonic saline group, while $FrH_2O\;and\;Fr_{Na}$ did not associated with the decrease in $C_{Li}$. 5) $C_{Li}$ of the rabbits received dextran solution fluctuated persistently and $Fr_{Na}\;and\;FrH_2O$ did not change in along with $C_{Li}$ although $Fr_{Na}$ had a tendency to be higher than that in hypertonic saline group. 6) From the above results it was concluded that: (a) In rabbits with normal body store of $Na^+$, the $Fr_{Na}$ of renal tubule beyond proximal tubule. calculated from $C_{Li}$ as a measure of inflow from proximal tubule is constant in spite of variations in $C_{Li}$. (b) The $FrH_2O$ calculated from $C_{Li}$ is dependent largely upon ADH rather than inflow from proximal tubule. (c) When there is a decrease in plasma $Na^+$ concentration or ineffective body fluid. $Li^+$ reabsorption may occur in the thick segnent of Henle's loop and hence the determination of $Fr_{Na}$ and $FrH_2O$ will not be easy one, but $Fr_{Na}$ is constant under the same experimental conditions.

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Development of High-performance Microwave Water Surface Current Meter for General Use to Extend the Applicable Velocity Range of Microwave Water Surface Current Meter on River Discharge Measurements (전자파표면유속계를 이용한 하천유량측정의 적용범위 확장을 위한 고성능 범용 전자파표면유속계의 개발)

  • Kim, Youngsung;Won, Nam-Il;Noh, Joonwoo;Park, Won-Cheol
    • Journal of Korea Water Resources Association
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    • v.48 no.8
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    • pp.613-623
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    • 2015
  • To overcome the difficulties of discharge measurements during flood season, MWSCM(micowave water surface current meter) which measures river surface velocities without contacting water has been applied in field work since its development. The existing version of MWSCM is for floods so that its applicability is low due to the short periods of floods. Therefore the renovative redesign of MWSCM to increase the applicability was conducted so that it can be applied to the discharge measurements during normal flows as well as flood ones by extending the measurable range of velocity. A newly developed high-performance MWSCM for general use can measure the velocity range of 0.03-20.0 m/s from flood flows to normal flows, whereas MWSCM for floods can measure the velocity range of 0.5-10.0 m/s. The improvement of antenna isolation between transmitter and receiver to block the inflow of transmitted singals to receiver and the improvement of phase noise of oscillator are necessary for detecting low velocity with MWSCM technology. Separate type antenna of transmitting and receiving signals is developed for isolation enhancement and phase locked loop synthesizer as an oscillator is applied to high-performance MWSCM for general use. Microwave frequency of 24 GHz is applied to the new MWSCM rather than 10 GHz to make the new MWSCM small and light for convenient use of it at fields. Improvement requests on MWSCM for floods-stable velocity measurement, self test, low power consumtion, and waterproof and dampproof-from the users of it has been reflected on the development of the new version of MWSCM.

Clinicopathologic Characteristics and Prognoses for Multicentric Occurrence and Intrahepatic Metastasis in Synchronous Multinodular Hepatocellular Carcinoma Patients

  • Li, Shi-Lai;Su, Ming;Peng, Tao;Xiao, Kai-Yin;Shang, Li-Ming;Xu, Bang-Hao;Su, Zhi-Xiong;Ye, Xin-Ping;Peng, Ning;Qin, Quan-Lin;Chen, De-Feng;Chen, Jie;Li, Le-Qun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.217-223
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    • 2013
  • Background: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and the outcomes for patients are still poor. It is important to determine the original type of synchronous multinodular HCC for preoperative assessment and the choice of treatment therapy as well as for the prediction of prognosis after treatment. Aims: To analyze clinicopathologic characteristics and prognoses in patients with multicentric occurrence (MO) and intrahepatic metastasis (IM) of synchronous multinodular hepatocellular carcinoma (HCC). Methods: The study group comprised 42 multinodular HCC patients with a total of 112 nodules. The control group comprised 20 HCC patients with 16 single nodular HCC cases and 4 HCC cases with a portal vein tumor emboli. The mitochondrial DNA (mtDNA) D-loop region was sequenced, and the patients of the study group were categorized as MO or IM based on the sequence variations. Univariate and multivariate analyses were used to determine the important clinicopathologic characteristics in the two groups. Results: In the study group, 20 cases were categorized as MO, and 22 as IM, whereas all 20 cases in the control group were characterized as IM. Several factors significantly differed between the IM and MO patients, including hepatitis B e antigen (HBeAg), cumulative tumor size, tumor nodule location, cirrhosis, portal vein and/or microvascular tumor embolus and the histological grade of the primary nodule. Multivariate analysis further demonstrated that cirrhosis and portal vein and/or microvascular tumor thrombus were independent factors differentiating between IM and MO patients. The tumor-free survival time of the MO subjects was significantly longer than that of the IM subjects ($25.7{\pm}4.8$ months vs. $8.9{\pm}3.1$ months, p=0.017). Similarly, the overall survival time of the MO subjects was longer ($31.6{\pm}5.3$ months vs. $15.4{\pm}3.4$ months, p=0.024). The multivariate analysis further demonstrated that the original type (p=0.035) and Child-Pugh grade (p<0.001) were independent predictors of tumor-free survival time. Cirrhosis (p=0.011), original type (p=0.034) and Child-Pugh grade (p<0.001) were independent predictors of overall survival time. Conclusions: HBeAg, cumulative tumor size, tumor nodule location, cirrhosis, portal vein and/or microvascular tumor embolus and histological grade of the primary nodule are important factors for differentiating IM and MO. MO HCC patients might have a favorable outcome compared with IM patients.

Influence of Column Aspect Ratio on the Hysteretic Behavior of Slab-Column Connection (슬래브-기둥 접합부의 이력거동에 대한 기둥 형상비의 영향)

  • Choi, Myung-Shin;Cho, In-Jung;Ahn, Jong-Mun;Shin, Sung-Woo
    • Journal of the Korea Concrete Institute
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    • v.19 no.4
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    • pp.515-525
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    • 2007
  • In this investigation, results of laboratory tests on four reinforced concrete flat plate interior connections with elongated rectangular column support which has been used widely in tall residential buildings are presented. The purpose of this study is to evaluate an effect of column aspect ratio (${\beta}_c={c_1}/{c_2}$=side length ratio of column section in the direction of lateral loading $(c_1)$ to the direction of perpendicular to $c_1$) on the hysteretic behavior under earthquake type loading. The aspect ratio of column section was taken as $0.5{\sim}3\;(c_1/c_2=1/2,\;1/1,\;2/1,\;3/1)$ and the column perimeter was held constant at 1200mm in order to achieve nominal vertical shear strength $(V_c)$ uniformly. Other design parameters such as flexural reinforcement ratio $(\rho)$ of the slab and concrete strength$(f_{ck})$ was kept constant as ${\rho}=1.0%$ and $f_{ck}=40MPa$, respectively. Gravity shear load $(V_g)$ was applied by 30 percent of nominal vertical shear strength $(0.3V_o)$ of the specimen. Experimental observations on punching failure pattern, peak lateral-load and story drift ratio at punching failure, stiffness degradation and energy dissipation in the hysteresis loop, and steel and concrete strain distributions near the column support were examined and discussed in accordance with different column aspect ratio. Eccentric shear stress model of ACI 318-05 was evaluated with experimental results. A fraction of transferring moment by shear and flexure in the design code was analyzed based on the test results.

Effect of Drainage Reusing Ratio on Growth and Yield of Summer-cultivated Paprika in Recycling Hydroponic Cultivation (순환식 수경재배에서 배액 재사용율이 여름작형 파프리카의 생육 및 수량에 미치는 영향)

  • Jang, Dong-cheol;Choi, Ki-Young;Kim, II-Seop
    • Journal of Bio-Environment Control
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    • v.26 no.1
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    • pp.7-12
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    • 2017
  • This experiment was conducted to analyze the effect of drainage reuse rate on the growth and fruiting of summer paprika in closed hydroponic cultivation. The experiment was carried out for 25 weeks from March to September 2015 with 0, 20, 30, 50% mixing ratio of waste nutrient solution using non - recycling hydroponic cultivation as a control. As a result, stem diameter of the test was different in the groups 1 and 2, but no difference showed as the group progressed more than 3 groups. L.A.I tended to decrease with increasing drainage mixing ratio. The number of nodes in the 50% reuse test group was 1.4 compared to the control group, but there was no significant difference. The number of harvested nodes was significantly different in the control group (11.1 nodes) and the 50% reuse test group (8.7 nodes), and the harvested nodes tended to decrease as the drainage was reused. The ratio of harvest was also the same as that of the harvesting node, and the control was the highest at 33.2% and the lowest at the 50% reuse test at 27.6%. Relative yields were reduced by 30%, 35% and 45% in the control group in the first group, and this tendency was also observed in the second and fourth groups. However, in the 3 and 5 groups, the production of 50% test group increased by 13% and 5%. The ratio of unmarketable fruit was increased 2%, 4%, 4%, and 7% in 0%, 20%, 30% and 50% reuse test, respectively. In conclusion, if the decrease in yield due to the decline in early growth is carefully managed, even if the imbalance of inorganic ions occurs after the mid-term growth, the growth of the crop will enter into a stable period and the re-use will not be worried about the growth and the yield decrease.

Binding Mode Analysis of Bacillus subtilis Obg with Ribosomal Protein L13 through Computational Docking Study

  • Lee, Yu-No;Bang, Woo-Young;Kim, Song-Mi;Lazar, Prettina;Bahk, Jeong-Dong;Lee, Keun-Woo
    • Interdisciplinary Bio Central
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    • v.1 no.1
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    • pp.3.1-3.6
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    • 2009
  • Introduction: GTPases known as translation factor play a vital role as ribosomal subunit assembly chaperone. The bacterial Obg proteins ($Spo{\underline{0B}}$-associated ${\underline{G}}TP$-binding protein) belong to the subfamily of P-loop GTPase proteins and now it is considered as one of the new target for antibacterial drug. The majority of bacterial Obgs have been commonly found to be associated with ribosome, implying that these proteins may play a fundamental role in ribosome assembly or maturation. In addition, one of the experimental evidences suggested that Bacillus subtilis Obg (BsObg) protein binds to the L13 ribosomal protein (BsL13) which is known to be one of the early assembly proteins of the 50S ribosomal subunit in Escherichia coli. In order to investigate binding mode between the BsObg and the BsL13, protein-protein docking simulation was carried out after generating 3D structure of the BsL13 structure using homology modeling method. Materials and Methods: Homology model structure of BsL13 was generated using the EcL13 crystal structure as a template. Protein-protein docking of BsObg protein with ribosomal protein BsL13 was performed by DOT, a macro-molecular docking software, in order to predict a reasonable binding mode. The solvated energy minimization calculation of the docked conformation was carried out to refine the structure. Results and Discussion: The possible binding conformation of BsL13 along with activated Obg fold in BsObg was predicted by computational docking study. The final structure is obtained from the solvated energy minimization. From the analysis, three important H-bond interactions between the Obg fold and the L13 were detected: Obg:Tyr27-L13:Glu32, Obg:Asn76-L13:Glu139, and Obg:Ala136-L13:Glu142. The interaction between the BsObg and BsL13 structures were also analyzed by electrostatic potential calculations to examine the interface surfaces. From the results, the key residues for hydrogen bonding and hydrophobic interaction between the two proteins were predicted. Conclusion and Prospects: In this study, we have focused on the binding mode of the BsObg protein with the ribosomal BsL13 protein. The interaction between the activated Obg and target protein was investigated with protein-protein docking calculations. The binding pattern can be further used as a base for structure-based drug design to find a novel antibacterial drug.

Research status of the development of genetically modified papaya (Carica papaya L.) and its biosafety assessment (GM 파파야 개발 및 생물안전성 평가 연구 동향)

  • Kim, Ho Bang;Lee, Yi;Kim, Chang-Gi
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.171-182
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    • 2018
  • Papaya (Carica papaya L.) is one of the crops widely planted in tropical and subtropical areas. The papaya fruit has low calories and are plentiful in vitamins A and C and in minerals. A major problem in papaya production is a plant disease caused by the papaya ringspot virus (PRSV). The first PRSV-resistant GM papaya expressing a PRSV coat protein gene was developed by USA scientists in 1992. The first commercial GM papaya cultivars derived from the event was approved by the US government in 1997. Development of transgenic papayas has been focused on vaccine production and limited agricultural traits, including insect and pathogen resistance, long shelf life, and aluminum and herbicide tolerance. Approximately 17 countries, including the USA and China, produced transgenic papayas and/or commercialized them, which provoked studies on biosafety assessment and development of GM-detection technologies. For the biosafety assessment of potential effects on human health, effects of long-term feeding to model animals have been studied in terms of toxicity and allergenicity. Studies on environmental safety assessment include influence on soil-microbial biodiversity and transfer to soil bacteria of GM selection markers. Many countries, such as Korea, the European Union, and Japan, that have strict regulations for GM crops have serious concerns about unintended introduction of GM cultivars and food commodities using unauthorized GM crops. Transgene- and/or GM event-specific molecular markers and technologies for genomics-based detection of unauthorized GM papaya have been developed and have resulted in the robust detection of GM papayas.