• Journal of Plant Biology
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• 제39권4호
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• pp.279-286
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• 1996

To know the changes of rbcL mRNA level by illumination, Northern hybridization analysis was performed with maize (Zea mays L.cv. Golden X Bantam). The average level of rbcL. mRNA in the light-grown shoots was 3.1 times higher than that of the dark-grown shoots after 6 to 10 growth days. The maximum difference of rbcL mRNA level between the dark-grown and the light-grown shoots was 5.1 folds. These results indicate that accumulation of rbcL mRNAin maize shoots is induced by light. Since the transcriptional DNA binding proteins and their cognate promoter elements, we carried out gel-retardation assays to elucidate the specific binding proteins on the rbcL promoter. It was found that plastid proteins of light-grown shoots bound to the R2 DNA fragment (-33 to -229) and R3 DNA fragment (-230 to -418 from ATG) of the rbcL promoter. From the results of competitive binding assays and heat or protease treatments, it was demonstrated that the bindings were sequence-specific DNA-protein interactions. Therefore, it could be concluded that the rbcL promoter region has at least two specific recognition sites for plastid proteins.

• Fisheries and Aquatic Sciences
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• 제15권1호
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• pp.43-47
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• 2012

Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.

• Journal of Animal Science and Technology
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• 제53권1호
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• pp.15-22
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• 2011

자돈의 생시 체중은 자돈의 이유 체중 및 생존율에도 크게 영향을 미친다. 또한 출하 시까지 돼지의 성장률 뿐만아니라 출하 일령 단축 및 출하 체중과도 밀접한 연관성이 있는 것으로 알려져 있다. 본 연구는 돼지 7번 염색체의 PGK 2 (phosphoglycerate kinase 2) 유전자의 promoter 영역 및 transcription 영역에 해당하는 DNA 염기서열을 유전체 구조분석을 통해 20개의 SNP (Single Nucleotide Polymorpism)를 발굴하였고, 발굴된 SNP 중 PGK 2 유전자의 전사 조절 영역 내 g.122 T>G 다형을 재래돼지와 Landrace를 이용한 $F_2$ 집단 268두 자돈의 성장 형질과의 연관성 분석을 실시한 결과, 생시 체중(p<0.01) 및 3주령 체중(p<0.001)에서 통계적으로 고도의 유의적 연관성이 있음을 확인 할 수 있었다. 따라서 본 연구를 통하여 확인된 PGK 2 유전자의 promoter 영역 내 성장 형질 관련 SNP는 건강한 자돈 및 종돈을 조기 선발하기 위한 유전자 marker로 활용 가능성을 제시하였다.

• Reproductive and Developmental Biology
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• 제37권3호
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• pp.91-96
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• 2013

Transgenic chickens have been spotlighted as an highly potent bioreactor for their fecundity, short generation time, and eggs associated with mass production of protein. In this study, we generated transgenic chickens exhibiting oviduct specific expression of human growth hormone fused to human transferrin for oral administration. Gene of the modified growth hormone located at downstream ovalbumin promoter (~3.6 kb) was introduced to stage X blastodermal cell employing retrovirus vector system. Several transgenic chickens were successfully generated. However, genomic analyses showed unexpected deletion within the transgene. The modification of the transgene seemed to occur during germ cell formation because the deletion was detected only from the sperm DNA of the G0 founder animal. There was no evidence of deletion in the somatic cell DNA samples of the same chicken. Consequently, same pattern of the deletion was confirmed in both somatic and germ cells of the G1 progeny.

• KSBB Journal
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• 제11권4호
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• pp.431-437
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• 1996

본 연구에서는 질산염(nitrate) 존재하에서 혐기 조건이 되면 최대로 발현되는 변형된 naT 프로모터 를 대장균내에서 단백질을 대량 생산하기 위한 발현 운반체로 쓰일 수 있는지를 조사하였다. 이를 위해 용존산소농도에 따라 naT 프로모터의 유도에 영향을 미치는 염색체 fnT 유전자가 발현되지 못하는 대장 균(ES200l)에 pBR322 플라스미드에 프로모터상 의 10 지역에서 특정부위돌연변이화에 의해 con­s sensus sequence로 바핀 변형된(modified) naT 프 로모터(pMW616)가 도입되어 있는 계(pMW616/ E ES2001 )가 이용되었다. 이 변형된 naT 프로모터의 하류에는 구조유전자(structural gene) 인 질산염 환 원효소대 신에 ${\beta}$-galactosidase를 발현하는 lacZ 유 전자가 클로닝되어 있다. 이 변형된 naT 프로모터의 유도에 대한 최적조건을 찾기 위해 변형된 naT 프로 모터를 유도시키는 방법, 변형된 naT 프로모터가 최 대로 유도되는 질산엽의 농도, 말현된 $\beta$galactosid a ase의 양 빛 변형된 naT 프로모터가 유도되는 특성 들이 조사되었다. 이에 대한 실험으로부터 다음과 같은 결론들이 얻어졌다; 이 변형된 naT 프로모터로 부터 ${\beta}$-galactosidase의 말현은 질산염의 농도에 의 해서는 크게 영향을 받지 않았다. 한편, 변형된 naT 프로모터로부터 ${\beta}$-galactosidase의 말현은 성장단계 에서 대장균을 호기상태에서 OD600이 약 2.27~ 될 때까지 키우다가 유도시 혐기상태로 만드는 것이 가 장 유리하였으며, 이 때 발현된 ${\beta}$-galactosidase의 비활성도는 약 13,000 Miller unit였다. 하지만, 이 변형된 naT 프로모터는 이켓을 유도시키기 전에도 발현된 ${\beta}$-galactosidase의 비활성도가 약 6,000 M Miller unit로 매우 높음으로 인하여 이 변형된 naT 프로모터는 원하는 단백질을 유도성보다는 구성적으 로 발현시키는데 더 적합한 프로모터였다.

• 한국가축번식학회지
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• 제17권4호
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• pp.375-383
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• 1994

The human growth hormone (hGH) gene uder the control of the rat $\beta$-casein promoter gene was designed to produce transgenic mouse expressed hGH gene in only mammary gland. One hundred seventy two eggs microinjected were transferred to the oviducts of pseudopregnants and 43 offspring were delivered. By Southern blotting hybridization, 3 were transgenic with rat $\beta$-casein/hGH gene. The copy numbers of three transgenic founder were 1, 5, and 15, respectively. A radioimmunoassay was developed to quantitate the amount of expression of the hGH gene in mammary gland of transgenic mice. The amount of hGH was 13.3ng/ml in the lactating milk of one transgenic line, showing predominantly higher than 3.0ng/ml in milk of control mice. Therefore, our findings suggested that $\beta$-casein promoter may induce the tissue specific expression of structural gene.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.431-438
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• 2015

Ever since the ban on antibiotic growth promoters (AGPs), the livestock death rate has increased owing to pathogenic bacterial infections. There is a need of developing AGP alternatives; however, the mechanisms by which AGP enhances livestock growth performance are not clearly understood. In this study, we fed 3-week-old swine for 9 weeks with and without AGPs containing chlortetracycline, sulfathiazole, and penicillin to investigate the effects of AGPs on swine gut microbiota. Microbial community analysis was done based on bacterial 16S rRNA genes using MiSeq. The use of AGP showed no growth promoting effect, but inhibited the growth of potential pathogens during the early growth stage. Our results showed the significant increase in species richness after the stabilization of gut microbiota during the post-weaning period (4-week-old). Moreover, the swine gut microbiota was divided into four clusters based on the distribution of operational taxonomic units, which was significantly correlated to the swine weight regardless of AGP treatments. Taxonomic abundance analysis indicated a negative correlation between host weight and the abundance of the family Prevotellaceae species, but showed positive correlation to the abundance of the family Spirochaetaceae, Clostridiaceae_1, and Peptostreptococcaeae species. Although no growth performance enhancement was observed, the use of AGP inhibited the potential pathogens in the early growth stage of swine. In addition, our results indicated the ecological succession of swine gut microbiota according to swine weight. Here, we present a characterization of swine gut microbiota with respect to the effects of AGPs on growth performance.

• Journal of Microbiology and Biotechnology
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• 제2권1호
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• pp.26-34
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• 1992

The effect of induction temperature on fermentation parameters has been investigated extensively using Escherichia coli M5248[pNKM21], a producer of recombinant human interleukin-2 (rhIL-2). In this recombinant microorganism, the gene expression of rhIL-2 is regulated by the cI857 repressor and $P_L$ promoter system. The recombinant fermentation parameters studied in this work include the cell growth, protein synthesis, cell viability, plasmid stability, $\beta$-lactamase activity, and rhIL-2 productivity. Interrelationships of such fermentation parameters have been analyzed through a quantitative assessment of the experimental data set obtained at eight different culture conditions. While the expression of rhIL-2 gene was repressed at culture temperatures below $34^\circ{C}$ with little effect on other fermentation parameters, under the conditions of rhIL-2 production $>(36~44^\circ{C})$ the cell growth, plasmid stability, and $\beta$-lactamase activity were, as induction temperature was increased, more profoundly reduced. Although the rhIL-2 content in the insoluble protein fraction was maximum at $40^\circ{C}$, total rhIL-2 production in the culture volume was found to be highest at the induction temperature of $36^\circ{C}$. This was in contrast to the previously known optimum induction temperature of the P$_{L}$ promoter system $>(40~42^\circ{C})$.Explanations for such a discrepancy have been proposed based on a product formation kinetics, and their implications have been discussed in detail.l.

• 한국양식학회:학술대회논문집
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• 한국양식학회 2006년도 수산관련학회 공동학술대회 발표요지집
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• pp.381-382
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• 2006

• Interdisciplinary Bio Central
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• 제3권3호
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• pp.10.1-10.8
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• 2011

Introduction: Hash functions are computer algorithms that protect information and secure transactions. In response to the NIST's "International Call for Hash Function", we developed a biological hash function using the computing capabilities of bacteria. We designed a DNA-based XOR logic gate that allows bacterial colonies arranged in a series on an agar plate to perform hash function calculations. Results and Discussion: In order to provide each colony with adequate time to process inputs and perform XOR logic, we designed and successfully demonstrated a system for time-delayed bacterial growth. Our system is based on the diffusion of ${\ss}$-lactamase, resulting in destruction of ampicillin. Our DNA-based XOR logic gate design is based on the op-position of two promoters. Our results showed that $P_{lux}$ and $P_{OmpC}$ functioned as expected individually, but $P_{lux}$ did not behave as expected in the XOR construct. Our data showed that, contrary to literature reports, the $P_{lux}$ promoter is bidirectional. In the absence of the 3OC6 inducer, the LuxR activator can bind to the $P_{lux}$ promoter and induce backwards transcription. Conclusion and Prospects: Our system of time delayed bacterial growth allows for the successive processing of a bacterial hash function, and is expected to have utility in other synthetic biology applications. While testing our DNA-based XOR logic gate, we uncovered a novel function of $P_{lux}$. In the absence of autoinducer 3OC6, LuxR binds to $P_{lux}$ and activates backwards transcription. This result advances basic research and has important implications for the widespread use of the $P_{lux}$ promoter.