• 한국미생물·생명공학회지
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• 제20권2호
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• pp.136-142
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• 1992

토양 분리균인 B.stearothermophilus chromosome의 유전자 은행으로부터 E.coli HB101 균주에 $\beta$-D-xylosidase 생산능력을 갖게하는 5.4Kb와 6.4Kb의 두 DNA 절편을 분리, pBR322에 크로닝하여 각각 pMG01과 pMG02의 재조합 플라스미드를 얻었다. 상기 두 B.stearothermophilus DNA 절편의 restriction map을 작성하고 이것을 기초로 하여 $\beta$-D-xylosidase 유전자인자의 위치를 확인함과 동시에 pUC18에 subcloning하여 각각 2.2kb와 1.0kb의 DNA 단편이 삽입된 $\beta$-D-xylosidase 양성의 pMG1와 pMG2를 분리하였다.

• 생명과학회지
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• 제15권6호
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• pp.928-934
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• 2005

ATF2는 c-Fos와 c-Jun과 같은 전사인자이며 이들은 루신지퍼 단백질이다. 루신지퍼 단백질은 동형이합체 혹은 이형이합체를 형성하며 promoter 영역에 결합하여 전사조절에 중요한 역할을 한다. 세포의 전사인자 ATF2는 ATF/CRE site에 결합하며 특히 선택적인 interfamily 이형이량체를 형성함으로써 전사 조절에 있어서 다양한 메카니즘을 제공할 수 있다. 본 연구에서는 ATF2 cDNA를 6xHis를 가진 expression vector에 subcloning하여 E.cnli BL2l에서 발현시켰다. 6xHis tag은 nickel-chelating chromatography를 가능하게 하였다 발현된 ATF2는 In vitro binding pull-down assay에서 동형이합체를 이를 뿐만 아니라 AP-1 그룹의 인자들과 선택적인 이형이합체를 형성함을 보여 주었다. BATF와는 강하게 결합하였으며 c-Jun과도 안정된 이합체를 형성하였다. 그러나 c-Fos와는 이합체를 형성하지 않으므로서 AP-1그룹 내에서도 이합체 형성이 선택적으로 이루어짐을 알 수 있다.

• Journal of Microbiology and Biotechnology
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• 제4권2호
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• pp.108-112
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• 1994

The genetic organization of the urease gene cluster from an alkalophilic Bacillus pasteurii was determined by subcloning and Tn5 transposon mutagenesis of a 10.7 kilobasepair cloned fragment. A region of DNA between 5.0 and 6.0 kb in length is necessary for urease activity. In vitro transcription-translation analysis of transposon insertion mutants of the cloned urease genes demonstrated that the major ($M_r$ 67,000) and minor ($M_r$ 20,000) structural peptides of urease are encoded at one end of the urease gene cluster and at least 3 additional polypeptides are encoded by adjacent DNA sequences.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.584-587
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• 2000

The invasion genes from Salmonella typhimurium were identified by the construction of a cosmid library and subcloning genes into a plasmid vector, pGEM-7Z. The 4.65 kb fragment of the invasion-conferring genomic region of the subclone, pSV6235 was sequenced in both direction. The three open reading frames, which were located at downstream of a promoter region, were designated as sir (Salmonella invasion region)A coding for the 36 amino acids, sirB coding for the 132 amino acids and sirC for the 82 amino acids, respectively. Interesingly, the genomic region of pSV6235 was highly homologous to Yersinia enterocolitica genomic DNA for a high pathogenicity island and Salmonella enteritidis insertion element IS1351 and IS200 DNA. These results show that there could be a significant relationship between S. typhimurium, Y. enterocolitica and S. enteritidis with respect to horizontal evolution process and acquisition of virulence determinants by means of transposon, plasmid or bacteriophage.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.527-530
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• 2000

중국산 야생키위로 total RNA를 추출하고 RT-PCR를 실시해서 증폭된 1.2kb fragment를 얻어 pGEM-T Easy에 cloning한 후 염기서열과 아미노산 서열을 비교 분석한 결과 actinidin gene인 것으로 분석되었다.

• Journal of Microbiology and Biotechnology
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• 제5권5호
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• pp.250-256
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• 1995

The ddh gene encoding meso-diaminopimelate (meso-DAP)-dehydrogenase (DDH) in Brevibacterium lactofermentum was isolated by complementation of the Escherichia coli dapD mutation. It was supposed from subcloning experiments and complementation tests that the evidence for DDH activity appeared in about 2.5 kb Xhol fragmented genome. The 2.5 kb Xhol fragment containing the ddh gene was sequenced, and an open reading frame of 960 bp encoding a polypeptide comprising 320 amino acids was found. Computer analysis indicated that the deduced amino acid of the B. lactofermentum ddh gene showed a high homology with that of the Corynebacterium glutamicum ddh gene.

• 한국미생물·생명공학회지
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• 제19권1호
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• pp.21-24
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• 1991

알칼리 내성 Bacillus sp. YA-14의 chromosomal DNA로부터 분리된 promoter를 subcloning하여 생화학적 특성을 조사하였다. B.subtilis와 promoter 공여균주, Bacillus sp. YA-14에서 promoter의 활성은 포자형성 초기단계에서 급격히 증가하였으며 1.0(w/v) glucose 첨가로 promoter 활성이 억제되었고 c-GMP에 의해 저해되었던 활성이 증가하였다.

• Journal of Microbiology and Biotechnology
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• 제1권4호
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• pp.251-255
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• 1991

A 16 kilobase (kb) HindIII fragment of alkalophilic Bacillus sp. YC-335 containing a gene for xylanase synthesis was inserted at the HindIII site of pBR322 and cloned in Escherichia coli HB101. After subcloning of recombinant plasmid pYS52, the 1.5 kb fragment was found to code for xylanase activity, and the hybrid plasmid was named pYS55. The DNA insert of the plasmid was subjected to restriction enzyme mapping, which showed that pYS55 had single site for PuvII and SstI in the 1.5 kb insert fragment. Southern hybridization analysis revealed that the cloned gene was hybridized with chromosomal DNA from alkalophilic Bacillus sp. YC-335. About 64% of the enzyme activity was observed in the extracellular and periplasmic space of E. coli HB10l carrying pYS55.

• 약학회지
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• 제35권1호
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• pp.22-29
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• 1991

pMB4 is a 2.4-kilobase plasmid of Staphylococcus aureus TR-1 that confers inducible resistance to the macrolide-lincosamide-streptogramin B(MLS) antibiotics. By subcloning studies, it was found that the MLS resistance determinant was located at 1.0Kb fragment between Sau3AI and TaqI sites. DNA sequence of the MLS resistant determinant, named ermC-4 was determined, and found to be highly homologous with that of ermC. Because the leader peptide sequence of ermC-4 was identical with that of ermC, the expression of the resistance gene is thought to be controlled by posttranscriptional attenuation in S. aureus TR-1.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.429-432
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• 1990

알카리 내성 Bacillus sp. YA-14의 chromosomal DNA로 부터 유래된 promoter를 subcloning하여 그 생화학적 특성과 포자형성과의 관계를 조사하였다. 알카리내성 Bacillus sp.와 B.subtilis에서 promoter의 활성은 포자가 형성되기 시작하는 단계에서 증가하기 시작했다. 또한 배지내 glucose가 1.0(w/v) 첨가시 promoter 활성이 억제되었다. 5가지의 spoO 유전자 중에서 3가지 유전자 (spoOB, spoOH, spoOJ) 산물이 promoter 활성에 필요하다.