• Biotechnology and Bioprocess Engineering:BBE
• /
• 제9권1호
• /
• pp.23-34
• /
• 2004

Escherichia coli transcription termination factor Rho catalyzes the unwinding of RNA/DNA duplex in reactions that are coupled to ATP binding and hydrolysis. Fluorescence stopped-flow methods using ATP and the fluorescent 2'(3')-O-( N-methylanthraniloyl) derivatives (mant-derivatives) of ATP and ADP were used to probe the kinetics of nucleotide binding to and dissociation from the Rho-RNA complex. Presteady state nucleotide binding kinetics provides evidence for the presence of negative cooperativity in nucleotide binding among the multiple nucleotide binding sites on Rho hexamer. The binding of the first nucleotide to the Rho-RNA complex occurs at a bimolecular rate of 3.6${\times}$10$\^$6/ M$\^$-1/ sec$\^$-1/ whereas the second nucleotide binds at a slower rate of 4.7${\times}$10$\^$5/ M$\^$-1/ sec$\^$-1/ at 18$^{\circ}C$, RNA complexed with Rho affects the kinetics of nucleotide interaction with the active sites through conformational changes to the Rho hexamer, allowing the incoming nucleotide to be more accessible to the sites. Adenine nucleotide binding and dissociation is more favorable when RNA is bound to Rho, whereas ATP binding and dissociation step in the absence of RNA occurs significantly slower, at a rate ∼70- and ∼40-fold slower than those observed with the Rho-RNA complex, respectively.

• 한국양식학회지
• /
• 제11권3호
• /
• pp.295-301
• /
• 1998

Since the end of the Pliocene, ancestral strains of Astyanax fasciatus have been accidently washed into different caves at the time of flooding and have lost their eyes and body pigments. Availability of this independently derived cave fish and their ancestral form within a single species provided a unique opportunity for studying the process of molecular evolution of the visual pigment gene. The nucleotide sequence comparisons of an ancestral river fish and two cave fish showed that nucleotide polymorphism of a green-like visual pigment gene between the eyed and blind form of A. fasciatus was much higher than that between the same blind form. Considering the number of nucleotide substitutions per nucleotide site and the direction of the nucleotide substitutions, more nucleotide substituions between the different forms of fish rater than the same one were probably due to more frequent mutations in the eyed river form. Nucleotide substitutions per site at the intron have been ocurred more than three times faster than those at the exon. This result indicates that the functional constraint has affected the green-like visual pigment gene of the blind cave fish although its eye sight is no longer required.

• 한국동물위생학회지
• /
• 제27권1호
• /
• pp.103-109
• /
• 2004

Babesia equi ema-1 5' intergenic(IG) nucleotide was PCR amplified and analyzed for restriction sites in order to identify a promoter region in this IG nucleotide sequence. B equi ema-1 5' IG specific primers identified a 1268 bp PCR product. The sequence had restriction sites for 34 restriction enzymes when analyzed by a computer program. Among them, 26 enzymes had only one restriction site, but the others had more than one sites. When four restriction enzymes (Bgll , HindⅢ, Kpn1 and BamH1) were treated to digest the 1268 bp nucleotide, they had restriction sites as expected by the computer program. Information of restriction sites in the 1268 bp IG nucleotide will be applied to select restriction enzymes for cloning the IG nucleotide to a vector.

• Journal of Nutrition and Health
• /
• 제31권4호
• /
• pp.679-686
• /
• 1998

This study was performed to determine whether the infusion of nucleosides and a nucleotide mixture directly ito intestinal lumen can induce a regenerative effect on impaired intestinal mucosa. The effects of massive small bowel resention and also total parenteral nutrition were induced by surgical creation of Thirty-Vella fistual in male Sprague-Dawley rats. The rats received saline solution (Control group) or nucleosides and a nucleotide mixture(lower concentration group(Nucl) or higher concentration group (Nuc2) every two days into the fistula. Mucosal protein, DNA , ornithine decarboxylase(ODC) activity, and morphometry were evaluated at 9 or 21 days postoperation in the fistual and also in the residual ileal segment. On the 9th day, mucosal protein, DNA content, and villous surface area in the fistula and also in the residual ileum increased in rats that received nucleosides and a nucleotide mixture of lower concentration (Nuc 1). On the 21 th day, there were no significant differences in intestinal mucosa between the control group and the lower level nucleoside nucleotide mixture-treated group. The fistula villous height of the higher nucleosides and a nucleotide mixture group was higher than in the control rats. Fistula mucosal ODC activities were not significantly different between groups although the mucosal ODC activity of the residual ileal segment was increased on the 9th day. Our data suggests that this animal model is suitable for studying the effect of dietary factors on intestinal mucosal growth and regeneration after villous stropy , differentiating direct effects of diet on the intestine from systemic effects. It is also suggested that external nucleosides and nucleotides have supportive effects on intestinal mucosal regeneration.

• 생명과학회지
• /
• 제14권4호
• /
• pp.608-613
• /
• 2004

Pyrimidine nucleoade N-ribosidase (pyrimidine 5'-nucleotide phosphoribo (deoxyribo) hydrolase/ pyrimidine 5'-nucleotide nucleosidase, EC 3.2.2.10)는 CMP와 UMP를 직접 분해하여 cytosine과 uracil를 생성하는 가수분해효소이다. 본 연구에서는 Pseudomonas oleovorans ATCC 8062의 생육과 pyrimidine nu-cleotide N-ribosidase생성에 미치는 탄소원과 질소원의 영향 및 생육인자 등에 대하여 검토했다. 효소생성의 최적배양조건은 2% fumarate, 1.5% peptone, 5% corn steep liquor (CSL)과 1% ammonium chloride의 배지조성(초기 pH 7.0)으로 $28^{\circ}C$, 48시간 진탕 배양이 양호했다. 효소의 활성은 생육이 최대에 도달하는 정지기 후기에 최대에 도달하며, 그 이후부터 급속히 불활성화 되었다. P. oleovorans의 pyrimidine nucleocde N-ribosidase는 UMP에 의하여 유도 생성되지 않으므로 구성효소이며, 내생효소였다.

• 한국국제농업개발학회지
• /
• 제23권5호
• /
• pp.546-551
• /
• 2011

국화과(Compositae)는 현화식물 중 세계에서 가장 넓게 분포하고, 쌍자엽식물 중 가장 진화된 식물분류군이며, 우리나라에는 약 300여종이 존재하는 것으로 알려져 있다. 구절초, 감국, 쑥, 쑥갓, 개미취, 참취, 곰취 등 국화과 식물들은 예로부터 민간에서 약용 및 식용 소재로써 다양하게 사용되어왔다. 본 연구는 국화 및 국화근연종 유용유전자원 선발을 통하여 육종 소재를 확대하고, 중간모본 및 신품종 육성기반을 구축하고자 DNA 마커시스템의 개발을 위해 수행되었다. 1. 화단국인 Smileball(Dendranthema grandiflorum) 품종을 사용하여 SSR-enriched library를 작성하였고, GS FLX 분석을 통해 18.83Mbp의 염기서열 결과를 얻었으며, read의 평균 길이는 280.06bp로 나타났다. 2. 단순반복염기서열(SSR) 부위를 포함하는 26,780개 clones 중 di-nucleotide motifs가 16,375개(61.5%)로 우세하였고, tri-nucleotide motifs(6,616개, 24.8%), tetra-nucleotide motifs(1,674개, 6.3%), penta-nucleotide motifs(1,283개, 4.8%), hexa-nucleotide motifs(693개, 2.6%) 순으로 나타났다. 3. 얻어진 di-nucleotide motifs들 중에서는, AC/CA class가 93.5%로 대부분이었고, tri-nucleotide motifs에서는 AAC class가 50.5%, tetra-nucleotide motifs는 ACGT class가 43.6%이고, penta-nucleotide motif에서는 AACGT class 27.2%이며, hexa-nucleotide motif에서는 ACGATG class 21.8%였다. 4. 얻어진 염기서열 결과를 토대로 다양한 motif를 갖는 100개의 SSR 마커를 제작하였고, 차후 이를 활용하여 국화 유전자원의 다형성 및 유전자형 분석을 통해 분자유전학적 다양성 및 집단의 구조분석이 가능하고, 국화의 분자육종기반 구축을 위한 유용한 도구가 될 것 이다.

• 한국식물병리학회지
• /
• 제14권6호
• /
• pp.612-613
• /
• 1998

The nucleotide sequence of hop stunt viroid HHSVd) Kh strain was sequenced by the reverse transcription and polymerase chain reaction. It consists of 296 nucleotides, and differs by one nucleotide deletion of cytosine at the position of 295 from the HSVd-K strain which consists of 297 nucletoides.

• Asian-Australasian Journal of Animal Sciences
• /
• 제19권7호
• /
• pp.922-926
• /
• 2006

A genetic polymorphism study on butyrophilin gene was carried out to explore variability of this gene and to estimate effects of such variability on milk quality traits in crossbred cattle. Polymorphism was unraveled by conducting Hae III PCR-RFLP of this gene. Three genotypes such as AA, BB and AB and two alleles namely A and B were observed in crossbred population. The frequencies of genotypes and alleles were 0.78, 0.17 and 0.04 for AA, AB and BB genotypes, respectively, and 0.87 and 0.13 for A and B alleles, respectively. The nucleotides, which have been substituted from allele A to B, were observed as C to G ($71^{st}$ nucleotide), C to T ($86^{th}$ nucleotide), A to T ($217^{th}$ nucleotide), G to A ($258^{th}$ nucleotide), A to C ($371^{st}$ nucleotide) and C to T ($377^{th}$ nucleotide). The nucleotide substitutions at $71^{st}$, $86^{th}$ and $377^{th}$ position of the fragment were found as silent mutations whereas nucleotide changes at $217^{th}$, $258^{th}$ and $371^{st}$ positions were detected as substitution of amino acid lysine with arginine, valine with isoleucine, and leucine with proline from allele A to B. The genotypes had significant effects ($p{\leq}0.05$) on total milk solid%, fat%, SNF%, while showing nonsignificant impact on total protein%. AA genotype produced highest average yield for all the traits.

• Asian-Australasian Journal of Animal Sciences
• /
• 제29권1호
• /
• pp.29-35
• /
• 2016

The aim of the present study was to investigate the effect of single nucleotide polymorphisms in the major histocompatibility complex (MHC) class II gene on resistance to Newcastle disease virus and body weight of the Thai indigenous chicken, Leung Hang Khao (Gallus gallus domesticus). Blood samples were collected for single nucleotide polymorphism analysis from 485 chickens. Polymerase chain reaction sequencing was used to classify single nucleotide polymorphisms of class II MHC. Body weights were measured at the ages of 3, 4, 5, and 7 months. Titres of Newcastle disease virus at 2 weeks to 7 months were determined and the correlation between body weight and titre was analysed. The association between single nucleotide polymorphisms and body weight and titre were analysed by a generalized linear model. Seven single nucleotide polymorphisms were identified: C125T, A126T, C209G, C242T, A243T, C244T, and A254T. Significant correlations between log titre and body weight were found at 2 and 4 weeks. Associations between single nucleotide polymorphisms and titre were found for C209G and A254T, and between all single nucleotide polymorphisms (except A243T) and body weight. The results showed that class II MHC is associated with both titre of Newcastle disease virus and body weight in Leung Hang Khao chickens. This is of concern because improved growth traits are the main goal of breeding selection. Moreover, the results suggested that MHC has a pleiotropic effect on the titre and growth performance. This mechanism should be investigated in a future study.

• 한국식품영양과학회지
• /
• 제26권1호
• /
• pp.130-136
• /
• 1997

250g내외의 횐쥐에게 Thiry-Vella fistula를 만든 후 수술과 비경구 영양 효과로 인하여 위축되는 소장의 누공에 각각 nucleosides/nucleotide혼합용액 (Nuc군)이나 glutamine(Gln군), glycine(Gly군)등을 투여하여 소장 점막의 위축 방지나 재생효과가 있는지를 알아보고 자 실험하여 다음과 같은 결과를 얻었다. Thiry-Vella fistula 시술 후 9일 동안 2일에 한 번씩 총 4회 소장 누공으로 주입한 이미노산들과 nucleosides/nucleotide 혼합물의 효과는 누공의 소장 점막에 미치는 직접적인 효과와 비누공 부분에 대한 간접적인 효과로 나누어 볼 수 있다. Nucleosides/nucleotide 혼합물은 소장 누공으로 주입되어 직접 접촉되는 부분의 소장 DNA함량을 유의적으로 증가 시켰으며 단백질 함량도 높은 수치를 보여 주었고 조직 검사 결과 소장 융모의 길이와 표면적을 증가시킨 것으로 나타났다. 그러나 ［$^3$H］ thymidine incorporation의 결과는 대조군과 유의적인 차이를 보이지 않았다. Glycine은 소장 단백질과 DNA에 있어서 대조군보다는 높고 Nuc군보다는 낮은 경향을 보여 주었으나 ［$^3$H］-thymidine incorporation은 누공과 비누공부분에서 모두 가장 높게 나타났으며 특히 비누공부분에서는 유의적으로 증가되어 있었다. Glutamine의 효과는 glycine 투여 효과와 비슷하거나 오히려 떨어지는 것으로 나타났다.