• 한국미생물·생명공학회지
• /
• 제14권2호
• /
• pp.181-186
• /
• 1986

대장균 염색체의 acetyl CoA carboxylase (fabE)영역 (염색체 지도상 72분 영역)의 유전자를 가진 결함형질도입 파아지를 분리했다 이 형질도입 파아지로부터 20 Md의 염색체 유래의 DNA를 분리하여 제한효소 지도를 작성했으며 이 영역에는 제한효소 Eco RI의 절단부위는 없었다. 형질도입 파아지 DNA의 제한효소 분해산물들은 pACYC 184 플라스미드 벡터에 재클로닝하여 fab E의 온도감수성 변이를 회복할 수 있는 수 종류의 플라스미드를 분리했다. 이들 플라스미드를 분석하여 fab E 유전자는 7, 4Md Bel II 단편상의 Hind III의 절단부위를 가진 3.4 Md Ban HI-Sal I 단편내에 존재함을 밝혔다.

• Journal of Microbiology and Biotechnology
• /
• 제13권6호
• /
• pp.976-983
• /
• 2003

Seven Escherichia coli cells defective with aerobic growth were isolated by the insertion of ${\lambda}placMu53$, a hybrid bacteriophage of ${\lambda}$ and Mu, which created a transcriptional fusion to lacZY. These insertion mutant cells were tested on an XG ($5-bromo-4-chloro-3-indolyl-{\beta}-D-galactopyranoside$) medium for anaerobic expression of lacZ by fusion to a promoter. The chromosomal DNA from these strains were digested by EcoRI, and the EcoRI fragments that contained the fused gene and lacZ sequence were identified by Southern hybridization, using lacZ containing plasmid as a probe. The EcoRI fragment from each strain was cloned and sequenced. The sequence data were compared with the GenBank database. The mutated gene of three strains, CYT4, CYT5, and OS11, was found to be identical, and it was nrdAB that encoded ribonucleoside diphosphate reductase. The gene nrdAB was at min 50.5 on the Escherichia coli linkage map and 2,348,084 on the physical map, and is involved in hemAe-related reduction-oxidation reaction. OS6 and OS14 mutant strains had insertion at min 8.3 and the mutated gene was hemB. The hemB encodes 5-aminolevulinate dehydratase or porphobilinogen synthase. The OS3 mutant had insertion in cydB at min 16.6. The cydAB encodes cytochrome d oxidase. In the case of OS1, the fusion was made with sucA, the E1 component of ${\alpha}-ketoglutarate$ dehydrogenase.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권3호
• /
• pp.309-312
• /
• 2004

We performed exon trapping in order to locate functional genes on chicken chromosomes (GGA) and to identify functional gene sequences from chicken cosmids. Sequence analysis of 100 clones revealed 17 putative exons, five of which were identified with known sequences in a gene database search: thymopoietin beta (TMPO), U5 snRNP-specific 40 kDa protein (HPRP8BP), dihydropyridine receptor alpha 1 subunit (CACNL1A3), cystein string protein (CPS) and C15orf4. We attempted to map the genes to chicken chromosomes by using FISH and linkage analysis. The chromosomal localizations were GGA1 (TMPO), GGA10 (C15orf4), GGA23 (HPRP8BP) and GGA28 (CPS) by FISH and linkage analysis, while that of CACNL1A3 was predicted to be on a microchromosome by FISH but not by linkage analysis. Comparative mapping analyses between chickens and humans for the genes revealed both known and new synteny. The syntenic conservation between GGA1 and human chromosome (HSA) 12q23 (TMPO) and between GGA10 and HSA15q25 (C15orf4), were consistent with a recent publication, while two new syntenies were observed between GGA28 and HSA20q13.3 in CPS and between GGA23 and HSA1p34-35 in HPRP8BP. The information of presently mapped genes can contribute as anchor markers based on functional genes and the construction of a comparative map.

• 대한두경부종양학회지
• /
• 제24권2호
• /
• pp.155-161
• /
• 2008

Head and neck squamous cell carcinoma(HNSCC) still has poor outcome, and laryngeal cancer is the most frequent subtype of HNSCC. Therefore, there is a need to develop novel treatments to improve the outcome of patients with HNSCC. It is critical to gain further understanding on the molecular and chromosomal alteration of HNSCC to identify novel therapeutic targets but genetic etiology of squamous cell carcinoma of the larynx is so complex that target genes have not yet been clearly identified. Array based CGH(array-CGH) allows investigation of general changes in target oncogenes and tumor suppressor genes, which should, in turn, lead to a better understanding of the cancer process. In this study, We used genomic wide array-CGH in tissue specimens to map genomic alterations found in laryngeal squamous cell carcinomas. As results, gains of MAP2, EPHA3, EVI1, LOC389174, NAALADL2, USP47, CTDP1, MASP1, AHRR, and KCNQ5, with losses of SRRM1L, ANKRD19, FLJ39303, ZNF141, DSCAM, GPR27, PROK2, ARPP-21, and B3GAT1 were observed frequently in laryngeal squamous cell carcinoma tissue specimens. These data about the patterns of genomic alterations could be a basic step for understanding more detailed genetic events in the carcinogenesis and also provide information for diagnosis and treatment in laryngeal squamous cell carcinoma. The high resolution of array-CGH combined with human genome database would give a chance to find out possible target genes which were gained or lost clones.

• Journal of Microbiology and Biotechnology
• /
• 제15권4호
• /
• pp.844-854
• /
• 2005

We constructed a defined physical and genetic map of H. pylori strain 51, previously isolated from a Korean patient with a duodenal ulcer, by combining a restriction analysis by pulse-field gel electrophoresis with the construction of a BAC library. A Notl-digest of H. pylori strain 51 genome yielded seven fragments, from which the genomic size was estimated to be 1,698$\pm$24 kb. The BAC library was constructed from 50 to 200 kb fragments of HindIII-digested genomic DNA. From 700 BAC clones, an ordered overlapping maxi-set of 82 BAC clones was assembled that covered the entire genome. The positions of 15 genes were localized in the strain 51 genome with 4-22 kb of resolution and were compared with their orthologues in strain 26695 and strain J99. The arrangement of the 15 genes was identical in strain 51 and strain J99, except for flaA and hpaA. The plasticity zone of strain 51, like that of strain J99, was located in the single region, and was shorter than those of strain 26695 and strain J99. The strain 51 plasticity zone consisted of ORFs common only to strain 51 and J99 or to strain 51 and 26695, as well as strain 51-specific ORFs. Three genetic translocations and/or inversions were found between orthologue ORFs in strain 51 and strain J99. These results show that the chromosomal organization of strain 51 differs from Western strains such as strain 26695 and strain J99.

• 한국작물학회지
• /
• 제49권6호
• /
• pp.533-538
• /
• 2004

화성벼와 O. minuta 간 여교잡을 통해 반복친인 화성벼와 출수기, 간장 등 여러 작물학적 특성에서 뚜렷한 차이를 보이는 염색체단편 치환계통 "WH79006"을 육성하였는데 화성벼와 WH79006의 차이는 WH79006에 이입된 O. minuta 염색체 단편에 의한 것이라고 할 수 있다. WH79006이 화성벼의 유전적 배경에 O. minuta의 어느 염색체 단편을 가지고 있는지를 검정하기 위해 SSR마커를 이용하였다. 벼 염색체에 골고루 분포한 294개의 SSR 마커를 검정한 결과 최소한 28개의 이입 단편을 검정하였는데 이들은 2번 염색체를 제외한 모든 염색체에 분포하였으며 전체 길이는 약 168cm이었다. 간장 관여 QTL을 탐색하기 위해 화성벼/WH79006 조합의 75개 $F_2$ 개체를 육성, 간장을 조사하였다. QTL분석 결과 6번 염색체의 RM225부근에서 간장에 관여하는 QTL cl6가 탐지되었다. cl6는 전체 표현형변이의 $9.6\%$를 설명하였으며 O. minute의 대립유전자가 간장을 크게 하는 방향으로 작용하였다. 이 결과는 O. minuta가 포복형임에도 불구하고 간장 조절 유전자들 중에는 간장을 크게 하는 방향으로 작용하는 유전자가 있음을 제시하고 있으며, 이 유전자들은 앞으로 QTL 분석을 통해 그 작용을 밝힐 예정이다. 추후교잡 집단을 이용 O. minuta 특이적 단편들이 어느 형질과 관련이 있는지를 밝힐 예정이다.

• 미생물학회지
• /
• 제23권2호
• /
• pp.138-146
• /
• 1985

포도당 이성화효소를 coding는 Bacillus licheniformis ATCC31667의 유전자를 Escherichia coli LE 392-6에 클로닝하였다. Bacillus lieheniformis 염색체 DNA를 분리하고 제한효소인 Pst I.HindIII, Sal 1, EcoR 1, BamH1으로 절단한 후 운반제 plasmid인 pBR332에 연결하고 포도당 이성화효소 negative인 E. coli LE 3926-6에 형질전환하였다. 이중 E채꺄 제한효소를 사용한 것만이 glucose isomerase positive로 전환되어 xylose를 유일 탄소원으로 하여 성장하였다. 이 제조합 plasmid를 제한효소로 처리하여 본 결과 4.1Kb의 Bacillus licheniformisdb전자가 옮겨 졌음을 확인했고 여기에 제한효소 HindII와 Puv II의 절단위치가 확인되어 제한요소 지도를 작정하였다. 이 재조합 plasmid pBGI6는 연속계대 10일 후에도 매우안정하게 유지되었다. 한편 포도당 이정화 효소의 안정을 측정하여 본 바 야생숙주에 비해 약 20배의 증가를 나타냈다.

• 정보처리학회논문지A
• /
• 제16A권3호
• /
• pp.143-152
• /
• 2009

최근 생물정보학 분야의 연구는 하나하나의 유전자를 연구하던 예전의 방법에서 유전자들간의 관계를 알아보는 연구들로 변해가고 있다. 이러한 유전자들 간의 연구 중 하나가 유전자 팀(gene team)을 연구하는 것이다. 유전자 팀이란 몇몇 염색체들 사이의 유전자들이 보존되어 있는 것을 말하며, 닫힌 영역 안에 보존되어 있는 유전자들의 집합으로 볼 수 있다. 이들은 진화과정을 거치면서, 유전자 팀 내의 유전자들의 위치나 그 종류가 변한다. 이러한 유전자 팀을 찾기 위해 많은 연구들이 이루어져왔다. 본 논문은 생물정보학 분야에서 많이 사용되는 계층적 클러스터링(hierarchical clustering)방법을 변형하여 전체 유전체(whole genome) 쌍내에서의 의미 있는 영역을 찾고, 영역 내에서 gene team을 찾을 수 있는 방법을 소개한다. 본 연구 방법을 이용하면, 복잡한 구조의 두 유전체 사이의 연관 유전자들이나 유사 영역들의 맵(map)을 단계별로 간략화 하여 나타낼 수 있다.

• 한국육종학회지
• /
• 제42권5호
• /
• pp.470-480
• /
• 2010

The objectives of this study were to identify QTLs for agronomic traits using introgression lines from a cross between a japonica weedy rice and a Tongil-type rice. A total of 75 introgression lines developed in the Tongil-type rice were characterized. A total of 368 introgressed segments including 285 homozygous and 83 heterozygous loci were detected on 12 chromosomes based on the genotypes of 136 SSR markers. Each of 75 introgression lines contained 0-9 homozygous and 0-8 heterozygous introgressed segments with an average of 5.8 segments per line. A total of 31 quantitative and 2 qualitative loci were identified for 14 agronomic traits and each QTL explained 4.1% to 76.6% of the phenotypic variance. Some QTLs were clustered in a few chromosomal regions. A first cluster was located near RM315 and RM472 on chromosome 1 with QTLs for 1,000 grain weight, culm length, grain width and thickness. Another cluster was detected with four QTLs for 1,000 grain weight, grain length, grain width and grain length/width ratio near the SSR marker RM249 on chromosome 5. Among the 31 QTLs, 9 (28.1%) Hapcheonaengmi3 alleles were beneficial in the Milyang23 background. ILs would be useful to confirm QTLs putatively detected in a primary mapping population for complex traits and serve as a starting point for map-based cloning of the QTLs. Additional backcrosses are being made to purify nearly isogenic lines (NILs) harboring a few favorable Hapcheonaengmi3 alleles in Milyang23 background.

• 한국육종학회지
• /
• 제40권1호
• /
• pp.1-7
• /
• 2008

An attempt was made to link rice embryo proteins to DNA sequences and to understand their functions. One hundred of the 700 spots detected on the embryo 2-DE gels were microsequenced. Of these, 28% of the embryo proteins were matched to DNA sequences with known functions, but 72% of the proteins were unknown in functions as previously reported (Woo et al. 2002). In addition, twenty-four protein spots with 100% of homology and nine with over 80% were matched to ESTs (expressed sequence tags) after expanding the amino acid sequences of the protein spots by Database searches using the available rice EST databases at the NCBI (http://www/ncbi.nlm.nih.gov/) and DDBJ (http://www.ddbj.nig.ac.jp/). The chromosomal location of some proteins were also obtained from the rice genetic map provided by Japanese Rice Genome Research Program (http://rgp.dna.affrc.go.jp). The DNA sequence databases including EST have been reported for rice (Oryza sativa L.) now provides whole or partial gene sequence, and recent advances in protein characterization allow the linking proteins to DNA sequences in the functional analysis. This work shows that proteome analysis could be a useful tool strategy to link sequence information and to functional genomics.