• Title/Summary/Keyword: genetic materials

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Production of ρ-Hydroxyacetophenone by Engineered Escherichia coli Heterologously Expressing 1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase

  • Wenmei Wu;Xiwei Yuan;Xin Gao;Chaoyang Tan;Shunxiang Li;Dehong Xu
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.467-475
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    • 2024
  • ρ-Hydroxyacetophenone is an important and versatile compound that has been widely used in medicine, cosmetics, new materials, and other fields. At present, there are two ways to obtain ρ-hydroxyacetophenone. One is to extract it from plants, such as Artemisia capillaris Thunb and Cynanchum otophyllum Schneid, and the other is to synthesize it by using chemical methods. Of these two methods, the second is the main one, although it has problems, such as flammable and explosive reagents, difficult separation of by-products, and harsh reaction conditions. To solve these issues, we adopted genetic engineering in this study to construct engineered Escherichia coli containing Hped gene or EbA309 gene. Whole-cell biotransformation was conducted under the same conditions to select the engineered E. coli with the higher activity. Orthogonal tests were conducted to determine the optimal biotransformation condition of the engineered E. coli. The results showed that the optimal condition was as follows: substrate concentration of 40 mmol/l, IPTG concentration of 0.1 mmol/l, an induction temperature of 25℃, and a transformation temperature of 35℃. Under this condition, the effects of transformation time on the ρ-hydroxyacetophenone concentration and cell growth were further studied. We found that as the transformation time extended, the ρ-hydroxyacetophenone concentration showed a gradually increasing trend. However, when the ρ-hydroxyacetophenone concentration increased to 1583.19 ± 44.34 mg/l in 24 h, cell growth was inhibited and then entered a plateau. In this research, we realized the synthesis of ρ-hydroxyacetophenone by biotransformation, and our findings lay a preliminary foundation for further improving and developing this method.

Expression of full and fragment-B of diphtheria toxin genes in Escherichia coli for generating of recombinant diphtheria vaccines

  • Shaimaa Abulmagd;Abd El-Nasser A. Khattab;Hamdallah Zedan
    • Clinical and Experimental Vaccine Research
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    • v.11 no.1
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    • pp.12-29
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    • 2022
  • Purpose: In the present study, whole diphtheria toxin (dt) and fragment B (dtb) genes from Corynebacterium diphtheriae Park William were cloned into Escherichia coli, the purified expressed proteins were evaluated for ultimately using as a candidate vaccine. Materials and Methods: The dt and dtb genes were isolated from bacterial strain ATCC (American Type Culture Collection) no. 13812. Plasmid pET29a+ was extracted by DNA-spin TM plasmid purification kit where genes were inserted using BamHI and HindIII-HF. Cloned pET29a+dt and pET29a+dtb plasmids were transformed into E. coli BL21(DE3)PlysS as expression host. The identity of the sequences was validated by blasting the sequence (BLASTn) against all the reported nucleotide sequences in the NCBI (National Center for Biotechnology Information) GenBank. Production of proteins in high yield by different types and parameters of fermentation to determine optimal conditions. Lastly, the purified concentrated rdtx and rdtb were injected to BALB/c mice and antibody titers were detected. Results: The genetic transformation of E. coli DH5α and E. coli BL21 with the pET-29a(+) carrying the dt and dtb genes was confirmed by colony polymerase chain reaction assay and were positive to grow on Luria-Bertani/kanamycin medium. The open reading frame of dt and dtb sequences consisted of 1,600 bp and 1,000 bp, were found to be 100% identical to dt and dtb sequence of C. diphtheriae (accession number KX702999.1 and KX702993.1) respectively. The optimal condition for high cell density is fed-batch fermentation production to express the rdtx and rdtb at 280 and 240 Lf/mL, dissolved oxygen was about 24% and 22% and the dry cell weight of bacteria was 2.41 g/L and 2.18 g/L, respectively. Conclusion: This study concluded with success in preparing genetically modified two strains for the production of a diphtheria vaccine, and to reach ideal production conditions to achieve the highest productivity.

Genetic Correlation of Carcass and Meat Production Traits with Hormones and Metabolic Components in Hawoo (가축의 혈청 호르몬 및 대사물질 농도와 도체 및 산육형질에 대한 유전상관에 관한 연구)

  • Jeon G. J.;Juong H. Y.;Cho K. H.;Kim M. J.;Kim I. C.;Kim J. B.
    • Journal of Embryo Transfer
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    • v.20 no.3
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    • pp.255-269
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    • 2005
  • This study was aimed to investigate genetic relationships, variables, and correlations between economic traits and metabolic materials in serum components according to bleeding periods and breeding locations for the castrated and not castrated Hanwoo cattle at National Livestock Research Institute. Analysis of variance for serum hormones and metabolic materials showed significant differences by breeding locations except for testosterone and globulin. Statistical differences for serum components were detected by birth year except for cortisol, total protein, globulin and creatinine, and by castration except for total protein and BUN. All the serum components were tended to have sire effects except for testosterone resulting in some degree of additive gene actions. Breeding locations showed statistical significances for carcass weight and back fat thickness, but not in carcass rate, KPH, live weight and transportation weight loss. Effects of breeding locations and castration were significant for all weight measurement periods except for 9 month and 6 month, respectively. A significant sire effect was observed in all weight measurements. Least squared means for concentration of serum components by breeding year, season and castration were not significant. High concentration of cortisol, creatinine and triglyceride and low concentration of IGF-1 and glucose were detected in castrated cattle. Concentration of testosterone with castrated cattle was $5.2\%$ corresponding to non castrated cattle. Estimation of heritabilities of serum components using a sire model with restricted maximum likelihood were ranged 0.07 to 0.58. High heritabilities were estimated for total protein, albumin, globulin, cortisol, creatinine and BUN were 0.53, 0.54, 0.42, 0.45, 0.58 and 0.54, respectively. Low heritabilities were estimated fur calcium, testosterone and IGF-1 for 0.07, 0.15 and 0.12, respectively. Heritabilities for carcass weight, back fat thickness, meat yield index, KPH, and IMF were estimated as 0.39, 0.45, 0.30 0.13, and 0.93. Heritabilities of weights on 18, 12, 9, 6, and 24 month were estimated as 0.78, 0.76, 0.62, 0.58 and 0.58. Estimated heritabilities for average daily gain on 6${\~}$2, 12${\~}$18, and 18${\~}$24 month were 0.80, 0.75 and 0.19, respectively.

Development of herbicide-tolerant Korean rapeseed (Brassica napus L.) cultivars (한국 고유의 품종을 이용한 제초제 저항성 유채 개발)

  • Kim, Hyo-Jin;Lee, Hye-Jin;Go, Young-Sam;Roh, Kyung-Hee;Lee, Young-Hwa;Jang, Young-Seok;Suh, Mi-Chung
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.319-326
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    • 2010
  • An interest in the production of seed-oil based fuel and raw materials, which comes from renewable plant sources, has been intrigued by the phenomenon of global warming and shortage of fossil fuels. Rapeseed (Brassica napus) is the most important oilseed crop, which produces seeds with 40% oil. It is desirable to develop genetically modified rapeseed producing oils, which can be easily converted to biodiesel. As an initial step for development of genetically modified rapeseed for the production of biofuels or bio-based materials, Korean rapeseed cultivars, Naehan, Youngsan, Tammi and Halla, were analyzed. Four Korean rapeseed cultivars produce 32 to 40% oil of seed dry weight, which is rich in oleic acid (more than 60 mole%). The cotyledonary petioles of rapeseed cultivar, Halla, were transformed using Agrobacterium tumefaciens strain GV3101, carrying the uidA gene encoding $\beta$-glucuronidase (GUS) as a reporter gene and the phosphinothricin acetyltransferase (PAT) gene as a selectable marker. The stable integration of PAT gene in the genome of transgenic rapeseeds was confirmed by PCR analysis. Expression of uidA gene in various rapeseed organs was determined by fluorometric assay and histochemical staining. Transformation efficiency of a Korean rapeseed Halla cultivar was 10.4%. Genetic inheritance of transgenes was confirmed in $T_2$ generation.

Molecular Characterization of Metallothionein Gene of the Korean Bitterling Acheilognathus signifer (Cyprinidae) (묵납자루 (Acheilognathus signifer; Cyprinidae) metallothionein 유전자의 클로닝 및 특징 분석)

  • Lee, Sang-Yoon;Bang, In-Chul;Nam, Yoon-Kwon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.10-20
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    • 2011
  • Genetic determinant for metallothionein (MT), a cysteine-rich protein playing essential roles in metal detoxification and homeostasis, was characterized in the Korean bitterling (Acheilognathus signifer, Cyprinidae), an endemic fish species. The full-length A. signifer MT (AsMT) cDNA (551 bp) is composed of a single open-reading frame (ORF) to encode a polypeptide of 60 amino acids containing 20 cysteine residues whose positions are conserved in most cypriniform MTs. At the genomic level, the AsMT (2,593 bp spanning the 5'-flanking region to the 3'-untranslated region) represented a conserved tripartite (three exons interrupted by two introns) structure with AT-rich introns. The upstream regulatory region (-1,914 bp from the ATG initiation codon) of AsMT displayed various sites and motifs for transcription factors involved in the metal-mediated regulation and stress/immune responses. The AsMT transcript was ubiquitously detected in various organs with variable expression levels, where the ovary and intestine showed the highest expression, while the heart and skeletal muscle represented the lowest level. During an exposure to copper (immersion in $0.5\;{\mu}M$ Cu for 48 h), the levels of AsMT transcripts were significantly elevated in the liver (more than 3.5-fold), moderately in the gill, kidney, and spleen (ranging from 1.5- to 2.5-fold), and barely in the brain and intestine. Results of this study could form a useful basis to explore the metal-related stress physiology of this endangered fish species.

Estimation of the quantitative trait loci associated with breaking and bending types lodging resistance in rice using chromosome segment substitution lines derived from a cross between Takanari and Koshihikari

  • Mulsanti, Indria Wahyu;Yamamoto, Toshio;Ueda, Tadamasa;Samadi, Ahmad Fahim;Adachi, Shunsuke;Hirasawa, Tadashi;Ookawa, Taiichiro
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.133-133
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    • 2017
  • Lodging is one of the important constraints in rice production. The lodging destroys the canopy structure, and sharply reduces the capacity of photosynthetic rate and dry matter production. In cereal crops, stem lodging can be classified into two types: stem breaking type and stem bending type. To improve stem lodging resistance, it is important to reveal strong culm traits of superior lodging resistant varieties. There are large varietal differences in parameters associated with the bending moment at breaking (M) and flexural rigidity (FR). The indica variety Takanari possesses large M due to its large section modulus (SM) despite of its small bending stress (BS), while Takanari also has large FR due to its large secondary moment of inertia (SMI) and Young's modulus (YM). To identify quantitative trait loci (QTLs) and the corresponding genes associated with the parameters for M ($=SM{\times}BS$) and FR ($=SM{\times}YM$) should enable to develop lodging resistant varieties, efficiently. In order to identify QTLs for cell wall materials such as cellulose, hemicellulose and lignin associated with BS and YM, a set of Chromosome Segment of Substitution Lines (CSSLs) consisted of 37 lines with chromosome segments of Koshihikari in the genetic background of Takanari were used. Takanari had large M and small BS as compared with Koshihikari. The QTLs for BS were estimated on chromosomes 3, 5, 6, 8, 9, 10, 11 and 12. Koshihikari alleles increased BS in these QTLs. Takanari had a large FR due to its large SMI and YM as compared with Koshihikari. The YM was increased by substitution of the Koshihikari chromosomal segments on chromosomes 2, 10 and 11. Other QTLs estimated on chromosomes 7 and 12 that Koshihikari alleles contributed to the decrease of YM. For lignin, only one major QTL for lignin density was detected on chromosome 11. Hollocellulose densities were increased by the substitution of Koshihikari segments on chromosomes 5 and 11. On the other hand, these were decreased on chromosomes 1 and 3 by substitution of Koshihikari segments. QTLs for cellulose density were estimated on chromosomes 1, 3 and 5 by substitution of Koshihikari segments. For hemicellulose, QTL on chromosome 3 showed that hemicellulose density decreased by the substitution of Koshihikari segment. However, hemicellulose densities on chromosomes 5, 8 and 11 showed the opposite effects. The QTLs for hemicellulose, cellulose, and hollocelulose densities identified on chromosome 5 overlapped with that for bending stress, indicating the positive effect of Koshihikari segment on increasing bending stress. These results suggest that some QTLs for the densities of cell wall materials contribute to increasing bending stress and Young's modulus, and could be utilized to improve the lodging resistance for both types of breaking and bending in rice varieties.

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The Optimum Conditions for Screening of Salt Resistant Lines Through Embryo Culture in Panax ginseng C.A. Meyer (인삼 염류내성 계통의 선발을 위한 배배양 배지조건)

  • Yang, Deok-Chun;Lee, Eun-Kyung;Kwon, Woo-Saeng
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.2
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    • pp.161-166
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    • 2003
  • Korean ginseng(Panax gmseng C.A. Meyer) is very difficult to obtain stable production of qualified ginseng roots because of variable stresses in soil environments. In environment stresses, soil condition is the most important factor, among which nutrients, especially inorganic materials such as N, P, K, Ca, Mg, Fe, etc., influence greatly on the ginseng growth. However, present ginseng field soils in Korea contain so much amount of such inorganic materials that a variety of remarkable disorders were noted in many ginseng plantations, resulting in decrease of qualitative ginseng production. Therefore, it is required to search for genetic resources and genes tolerant to salt stress for the development of ginseng cultivars. Selection of stress-tolerant ginseng lines in fields is very difficult because it is almost impossible to control properly the environmental conditions of soil. On the contrary, it can be studied with ease to search for stress-tolerant ginseng lines through in vitro culture because of easy manipulation of stress conditions. Murashige & Skoog(MS) media with 2.5 folds of $KNO_3,\;NH_4NO_3,\;MgSO_4\;7H_2O,\;KH_2PO_4,\;and\;CaCl_2\;2H_2O$ was established for the selection of ginseng lines tolerant to salt stress under the embryo culture.

Scab Resistance of Some Pear Genetic Resources and Inter Specific Hybrid Seedlings in Field Condition (포장조건에서 주요 배 유전자원과 종간교배 집단의 검은별무늬병 저항성)

  • Kim, Yoon-Kyeong;Jeong, Hae-Won;Won, Kyung-Ho;Kang, Sam-Seok;Lee, Ug-Yong;Han, Tae-Ho
    • Korean Journal of Plant Resources
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    • v.32 no.4
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    • pp.296-302
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    • 2019
  • This study was accomplished to select breeding materials among the pear germplasm 6 species 43 accessions and check out the expression of scab incidence of inter specific hybrids of 5 crosses 609 seedlings for scab resistance breeding. Fruitlets scab incidence of P. pyrifolia, P. bretschneideri and P. ussuriensis were 59.5, 73.4 and 63.0% respectively, and also fruitlets scab incidence of interspecific hybrid cultivars between P. pyrifolia and P. bretschneideri were 77.7%. However, fruitlets scab incidence of P. communis and interspecific hybrid accessions between P. pyrifolia and P. communis were significantly less than 15%. As a result of the Duncan test, scab resistance of pear accessions were classified into two groups: P. communis and their hybrid were resistance and P. bretschneideri, P. pyrifolia, P. ussuriensis, and among their hybrid were susceptible. Also, it showed the same trend in the seedlings. Scab incidence of 'Seolwon' ${\times}$ 'Chuwhangbae' and 'Supergold' ${\times}$ 'Mantienhong' in P. pyrifolia seedlings showed more than 80%, while that of interspecific hybrid between P. pyrifolia and P. communis showed about 2%. It showed scab resistance of P. communis transferred to hybrid seedlings and expressed to be dominant in their seedlings. Namely, P. communis was good breeding materials for scab resistance.

Characteristics and Genesis of Terrace Soils in Yeongnam Area -IV: Soil micromorphological features (영남지역(嶺南地域)에 분포(分布)된 단구지토양(段丘地土壤)의 특성(特性)과 생성연구(生成硏究) -제(第) 4 보(報) : 토양미세형태학적(土壤微細形態學的) 특성(特性))

  • Jung, Yeun-Tae;Shin, Jae-Sung;No, Yeong-Pal
    • Korean Journal of Soil Science and Fertilizer
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    • v.19 no.3
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    • pp.205-210
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    • 1986
  • A soil micromorphological study of clayey terrace soils in Yeongnam area was carried out to investigate the soil genetic features and to distinguish the soil parent materials. Dominant plasmic fabrics in the terrace soils were vosepic, mosepic, insepic, and masepic of sepic fabrics but the Geugrag and Upyeong in Yeongil sequence have an asepic fabrics. Embedded grain cutans in the terrace soils were characterized by having porphyroskelic related distribution. Among pedological features of the B horizons of the terrace soils, the argillic cutans of ferriargillans and glaebules of manganiferrous concretions were the dominant in mineralogical composition, and the papules are abundant in the most terrace soils indicating that the soils are generally old. The substrata of the Bancheon in Yeongcheon and Upyeong in Yeongil area were so different from the terrace deposits in plasmic fabrics that these soils were believed to be composed of bisequum profiles. Because of having the free grain cutans, it was considered that the substrata of Bancheon in Yeongcheon and Upyeong in Yeongil area might have a period of soil formation before deposition of the terrace materials.

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Genome Wide Association Study for Phytophthora sojae Resistance with the Two Races Collected from Main Soybean Production Area in Korea with 210 Soybean Natural Population

  • Beom-Kyu Kang;Su-Vin Heo;Ji-Hee Park;Jeong-Hyun Seo;Man-Soo Choi;Jun-Hoi Kim;Jae-Bok Hwang;Ji-Yeon Ko;Yun-Woo Jang;Young-Nam Yun;Choon-Song Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.202-202
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    • 2022
  • Recently days, soybean production in paddy field is increasing, from 4,422 ha in 2016 to 10,658 ha in 2021 in Korea. It is easy for Phytophthora stem and root rot (PSR) occurring in paddy field condition, when it is poorly drained soils with a high clay content, and temporary flooding and ponding. Therefore PSR resistant soybean cultivar is required. The objective of this study is to identify QTL region and candidate genes relating to PSR resistance of the race in main soybean cultivation area in Korea. 210 soybean materials including cultivars and germplasm were used for inoculation and genome-wide association study (GWAS). Inoculation was conducted using stem-scar method with 2 replications in 2-year for the race 3053 from Kimje and 3617 from Andong. 210 materials were genotyped with Soya SNP 180K chip, and structure analysis and association mapping were conducted with QTLMAX V2. The results of inoculation showed that survival ratio ranged from 0% to 96.7% and mean 9.7% for 3053 and ranged from 0% to 100% and mean 7.6% for 3617. Structure analysis showed linkage disequillibrium (LD) was decayed below r2=0.5 at 335kb of SNP distance. Significant SNPs (LOD>7.0) were identified in Chr 1, 2, 3, 4, 5, 11, 14, 15 for 3053 and Chr 1, 2, 3, 7, 10, 14 for 3617. Especially, LD blocks (AX-90455181;15,056,628bp~AX-90475572;15,298,872bp) in Chr 2 for 3053 and 3067 were duplicated. 29 genes were identified on these genetic regions including Glyma.02gl47000 relating to ribosome recycling factor and defense response to fungus in Soybase.

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