• 한국가금학회지
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• 제38권3호
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• pp.191-195
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• 2011

The objectives of this study are to identify specific functional genes which are related with growth and protein structure of the pectoral muscle of Korean native chicken. Pectoral muscle was isolated from three Korean native chickens (KNC, red brown, 12 months old, 2.41 ${\pm}$ 0.24 kg) and three Cornish chickens (16 month old, 2.76 ${\pm}$ 3.0 kg). The subtraction cDNA library was prepared in PCR4 Blunt-TOPO vector. The DNA sequence homology was compared with other breeds and species in GenBank. A clone NDS-81 was found to be unique for the DNA sequence homology with UBX family. Their partial sequence has high homology (98%) with chicken UBX domain D. Chicken UBX domain has chicken (93%), cattle (68%), dog (67%), mouse (64%) and, human (63%) nucleotide sequence homology. Several regions were mutated from T in chicken to C or G in the NDS-81 clone. The first site is LAD in chicken, but it was expressed as (L)RM in clone NDS-81. In this site, amino acids were changed from Ala to Arg, and from Asp to Met. The second site was changed from ER (Arg) in chicken to ED (Asp) in clone NDS-81. They are both containing functional side chains and play an important role in binding other proteins. Therefore, the clone NDS-81 could be a different candidate gene for the UBX family gene and could related with pectoral muscle structure of Korean native chicken.

• 한국동물학회지
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• 제35권2호
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• pp.203-210
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• 1992

Hsp70, a 70 kDa protein, is the maior protein expressed when cells are heat-shocked. A cDNA library from mouse ID13 cells was screened with the human hsp70 gene as a probe, and a positive clone was obtained. The positive clone was subcloned into puc19 and the precise restriction was obtained. The CDNA was sequenced by the Sanger's dideoxv termination method. Single open reading frame that codes for a protein of 70 kDa was found. The DNA sequence of the cloned mouse DNA shows great homology (66-90%) with other mouse hsp70 genes and somewhat less homology (50",) with E. coli hsp70 gene (dnak). With the exception of one amino acid, the protein sequence deduced from the CDNA is identical to the mouse that shock cognate protein 70 (hsc70) that is constitutivelv expressed at normal temperature. The result suggests that the cloned CDNA encodes a hsc70 family rather than a heatinducible family.mily.

• BMB Reports
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• 제51권9호
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• pp.437-443
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• 2018

Non-homologous end joining (NHEJ), and to a lesser extent, the error-free pathway known as homology-directed repair (HDR) are cellular mechanisms for recovery from double-strand DNA breaks (DSB) induced by RNA-guided programmable nuclease CRISPR/Cas. Since NHEJ is equivalent to using a duck tape to stick two pieces of metals together, the outcome of this repair mechanism is prone to error. Any out-of-frame mutations or premature stop codons resulting from NHEJ repair mechanism are extremely handy for loss-of-function studies. Substitution of a mutation on the genome with the correct exogenous repair DNA requires coordination via an error-free HDR, for targeted transgenesis. However, several practical limitations exist in harnessing the potential of HDR to replace a faulty mutation for therapeutic purposes in all cell types and more so in somatic cells. In germ cells after the DSB, copying occurs from the homologous chromosome, which increases the chances of incorporation of exogenous DNA with some degree of homology into the genome compared with somatic cells where copying from the identical sister chromatid is always preferred. This review summarizes several strategies that have been implemented to increase the frequency of HDR with a focus on somatic cells. It also highlights the limitations of this technology in gene therapy and suggests specific solutions to circumvent those barriers.

• 한국식물병리학회지
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• 제11권1호
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• pp.73-79
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• 1995

Complementary DNAs (cDNAs) to the coat protein gene of an ordinary Korean strain of potato virus Y (PVY-OK) isolated from potato (cv. Superior) were synthesized and cloned into a plasmid pUC119 and sequenced. The RNA of the virus propagated in tobacco (Nicotinaa sylvestris) was extracted by the method of phenol extraction. The first strand of cDNAs to the coat protein penomic RNA of the virus was made by Moloney murine leukemia virus reverse transcriptase. The cDNA were synthesized and amplified by the method of polymerase chain reaction (PCR) using a pair of oligonucleotide primers. PVYCP3P and PVYCP3M. The size of cDNAs inserted in pUC119 plasmid was estimated as about 840 bp upon agarose gel electrophoresis. Double stranded cDNAs were transformed into the competent cell of E. coli JM109. Sequence analysis of cDNAs was conducted by the dideoxynucleotide chain termination method. Homology of cDNAs of the PVY-OK coat protein genomic RNA with those of PVY-O (Japan), PVY-T (Japan), PVY-TH (Japan), PVYN (The Netherlands),and PVYY (France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. Homology at the amino acid level turned out to the be 97.4%, 92.5%, 92.9%, 92.9% and 98.5%, respectively.

• 한국미생물·생명공학회지
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• 제4권4호
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• pp.166-178
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• 1976

서로 다른 11주의 Bacillus coagulans와 13종의 Bacillus 속 14주를 deoxyribonucleic acid(DNA)-DNA hybridization method에 의해서 분류학적인 연구를 하였다. 사용한 B. coagulans 11주중 6주는 흙에서(일본 오사까교외) 분리했고, 나머지 5주는 ATCC, IFO에서 authentic strains을 얻어서 사용했다. 사용된 B. coagulans는 Bergey's Manual(8th ed)에 의거 Gordon씨들의 방법으로 동정한 결과 B. coagulans로서 확인되었다. 이렇게 동정된 B. coagulans을 분자 생물학적 차원에서 지금까지의 Conventional taxonomic study와의 관계를 연구하기 위해서 사용한 11주의 B. coagulans중 ATCC 7070을 $^3$H labeled input 즉 standard로 해서 사용했을 때 B. coagulans 내의 intraspecific DNA homology indexes는 76% 이상으로 나타났다. 이와같은 발견은 Bergey's Manual에 의거한 conventional taxonomic study의 결과와 잘 일치하고 있었음으로 새로 분리한 6주와 authentic sources로부터 받은 5주는 같은 group의 B. coagulans라는 사실을 입증해 주었다. 그리고 B. coagulans와 다른 species의 Bacillus 속 즉 B. pumilus(168), B. licheniformis (IFO 12107), B. pumilus(IFO12110), B. firmus(ATCC 14575), B. lentus(ATCC 10840), B. circulans(ATCC 4513), B.macelans(ATCC 8244), B.polymyxa, (ATCC 842), B.sphaericus(ATCC 14577), B.brevis(ATCC 8246, IFO 12334), B.laterosporus(ATCC 64), B. pantothenticus(ATCC 14576) interspecific DNA homology indexes가 각각 2～4%을 보임으로써 B. coagulans는 molecular level면에서 이들 Bacillus 속과는 상동성 관계가 적음을 나타내었다. 반면에 B. coagulans(ATCC 7050)와 E. coli(F-12)와의 상동성은 1%이하였다.

• 한국균학회지
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• 제26권3호통권86호
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• pp.314-320
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• 1998

mRNA의 differential display방법에 의해 여름느타리 자실체의 상처 또는 자외선 처리시 발현되는 약 0.4kb의 cDNA 단편을 분리하였다. 이 cDNA 단편의 염기서열 분석결과 세포분열 촉진에 관여하는 cdc2-related protein kinase gene과 상당부분 유사성을 보였으며 RT-PCR 방법을 이용한 분리 유전자의 발현 실험을 통해 이 유전자가 정상 생장 환경에서는 갓, 대, 균사 등 모든 부위에서 기본적인 발현상태를 유지하고 있으며 기계적 상처 또는 자외선 처리에 의해 그 발현이 증폭됨을 확인하였다. 이러한 결과를 통해 향후 분리된 유전자의 연구를 통한 버섯 병 방어 관련 신호 전달 체계 분석에 대한 가능성을 검토해 보았다.

• 생명과학회지
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• 제19권11호
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• pp.1522-1528
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• 2009

알긴산을 분해하기 위하여 갈조류로부터 분해활성이 있는 해양미생물을 분리하였다. 분리된 균주의 16S ribosomal DNA를 분석한 결과 이전에 보고했던 ALG-5 균주와 비슷한 Streptomyces sp.에 속하는 것으로 나타났다. 상동성이 있는 염기서열로 고안한 특이적인 primer로 PCR을 행함로서 Streptomyces sp. M3의 새로운 alginate lyase 유전자를 클로닝하였다. M3 alginate lyase의 예상 아미노산 서열에는 N-terminal 영역에 YXRSELREM 서열과 C-terimnal 영역에 YFKAGXYXQ 서열이 보존되어 있었다. M3 alginate lyase 단백질의 homology model은 Corynebacterium sp. ALY-1으로부터 얻은 단백질인 alyPG와 같이 $\beta$-jelly roll fold를 main domain으로 가지고 있음이 나타났다. M3 alginate lyase 유전자를 가지는 재조합 E. coli의 세포균질액은 polymannuronate block보다는 polyguluronate block에 대하여 높은 분해력을 가지고 있었다. 아미노산 서열 다중정열 및 homology modeling으로부터 얻은 결과는 M3 alginate lyase가 Family PL-7으로 분류될 수 있음을 말해 준다.

• Fisheries and Aquatic Sciences
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• 제4권4호
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• pp.192-196
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• 2001

Proopiomelanocortin (POMC) plays an essential role in the disease resistance system and is the precursor protein of biologically active peptides such as adrenocorticotropin (ACTH), $\alpha-melanocyte-stimulating$ hormone $(\alpha- MSH)$, $(\beta-melanocyte-stimulation hormone\;(\beta- MSH)$ and $\beta-endorphin$. We have isolated and sequenced two different forms of POMC cDNA, POMC-I and POMC-II, from a pituitary cDNA library of flounder. POMC-I cDNA consisted of 956 bp corresponding to deduced amino acids of 216 residues and POMC-II cDNA was 982 bp in length corresponding to 194 amino acids, respectively. The results of deduced amino acids analysis of the clones showed high sequence homology with previously reported POMCs amino acid sequences from various species. The homology between flounder POMC-I and -II is$57\%$ identity. We also constructed a phylogenetic tree based on POMC amino acid sequences.

• 한국식물병리학회지
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• 제13권2호
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• pp.113-117
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• 1997

Low moleuclar weight (LMW) RNAs were isolated form Korean peonies which expressed symptoms of stunt and epinasty. The LMW plant RNAs were purified by Qiagen column chromatography which could separate viroid specific nucleic acid at differential salt concentration. After the inoculation of the purified RNAs from the peonies, the inoculated tomatoes (cv. Rutgers) expressed the symptoms of stunt and epinasty. Also the same molecular weight RNAs with viroid-like RNAs were isolated from the inoculated tomatoes. Double-stranded cDNA were synthesized by the methods of reverse transcription (RT) and polymerase chain reaction (PCR) with the purified RNA and primers. The same cDNAs associated with viroid-like RNAs wre cloned from the inoculated tomatoes. The cDNA has been sequenced and its 375-nucleotides were arranged into secondary structure. The cloned cDNA showed 47~54% homology compared with other viroids. The sequence homology of the cloned cDNA were partially high with plant genomic RNAs.

• 생명과학회지
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• 제12권4호
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• pp.427-432
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• 2002

잇바디돌김(Porptyra dentata)을 대상으로 핵의 18S ribo-somal RNA를 지령하는 유전자 즉 18S rDNA 유전자를 증폭하고, 염기서열분석을 수행하였다. 전체 18S rDNA의 exon 영역 크기는 1822 bp, intron 영역의 크기는 512 bp였다. 이들 exon과 intron 영역의 G+C함량은 각각 49%와 55%씩 나타내었다. 일본산 잇바디돌김(CenBank accession number: AB013183)의 exon 영역과의 비교에서 상동성이 97.1%에 도달하였다. 568번과 569번 염기사이의 upstream에 위치하는 intron 영 역에서는 AB013183과 52.1%의 상동성을 보였다.