• 한국생물공학회:학술대회논문집
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• 2000.04a
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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.

• Mycobiology
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• v.28 no.4
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• pp.171-176
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• 2000

We have cloned a ${\alpha}-COP$ homolog, ancop, from Aspergillus nidulans by colony hybridization of chromosome specific library using ${\alpha}-COP$ homologous fragment as a probe. The probe DNA was amplified with degenerated primers designed by comparison of conserved region of the amino acid sequences of Saccharomyces cerevisiae ${\alpha}-COP$, Homo sapiens HEP-COP, and Drosophila melanogaster ${\alpha}-COP$. Full length cDNA clone was also amplified by RT-PCR. Comparison of genomic DNA sequence with cDNA sequence obtained by RT-PCR revealed 7 introns. Amino acid sequence similarity search of the anCop with other ${\alpha}-COPs$ gave an overall identity of 52% with S. cerevisiae, 47% with human and bovine, 45% with Drosophila and Arabidopsis. In upstream region from the transcription start site, a putative TATA and CAAT motif were also identified.

• Asian-Australasian Journal of Animal Sciences
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• v.6 no.4
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• pp.541-547
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• 1993

A phage clone containing the partial ${\alpha}_{S1}$-casein gene was isolated from a bovine genomic library by using mixed probes of ovine ${\alpha}_{S1}$-, ${\beta}$- and ${\kappa}$-casein cDNAs. Restriction enzyme mapping analysis for 14.6 kb revealed that the map was in conflict with the report of Meade et al. (1990), especially in the 3'-end fragment. Sequence analysis of 12.6 kb revealed a high AT/GC ratio (1.64); we have identified eight exon sequences according to the bovine ${\alpha}_{S1}$-casein cDNA sequence. The same exon/intron splice junction sequence was observed between these exons. We suggest that the bovine ${\alpha}_{S1}$-casein gene night contain a minimum of 18 exons and the full length is approximately 18-19 kb.

• Journal of Plant Biology
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• v.32 no.2
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• pp.67-78
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• 1989

Hybridization of DNA isolated from leaves of Russet Burbank potato with tomato cDNA as a probe revealed the presence of about ten inhibitor 1 genes in the genome. Screening of a genomic library of Russet Burbank potato resulted in isolation of seven different genomic clones carrying inhibitor I genes. One of the genomic clones, clone 2, contained two EcoRI fragments of 3.4 and 1.8 kb in size, respectively, which were hybridized with the probe. The nucleotide sequence of parts of the hybridizing EcoRI fragments revealed that they contain a complete gene which codes for an open reading frame of 107 amino acids. It is interrupted by two intervening sequences of 502 and 493 bp, situated at the positions of codons 17 and 43, respectively, of the open reading frame. Putative regulatory sequences, TATAAA and CCACT, were found at the 5' flanking region. In addition, a copy of a 100 bp repeat found at a tomato inhibitor I gene was identified.

• Journal of Plant Biology
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• v.32 no.2
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• pp.79-87
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• 1989

Southern hybridization of genomic DNAs with radioactively labeled cDNA of tomato proteinase inhibitor II revealed that proteinase inhibitor II proteins in potato plants are encoded by a family of about 10 related sequences. Screening of potato EcoRI genomic library with the cDNA resulted in isolation of 13 recombinant phage clones which carry 3 different genomic regions. Of these clones, clones 8, 18, and 39 were subjected to restriction mapping and subcloning. Further characterization of the subclones of clones 8, 18 and 39 indicated that two inhibitor II genes are present on a 8.0 kb EcoRI fragment of clone 8, one on 3.3 and 0.8 kb EcoRI fragments of clone 18 and two genes on a 13.5 kb EcoRI fragment of clone 39.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.21-26
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• 2010

As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

• Journal of Microbiology
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• v.38 no.4
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• pp.230-238
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• 2000

Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• Journal of Microbiology
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• v.43 no.6
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• pp.523-528
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• 2005

Using the genomic library constructed at the downstream of the niiA promoter, which induces the over-expression of an inserted DNA fragment, we have attempted to screen the genes affecting growth or development by over-expression. The wild-type strain was transformed using the AMA-niiA(p) library and cultured on 1.2 M sorbitol media, in which asexual sporulation is induced, but sexual development is repressed. Over 100,000 strains transformed to $pyrG^+$ were analyzed with regard to any changes in phenotype. Consequently, seven strains were isolated for further analyses. These strains were designated NOT [niiA(p) over-expression transformants] stains. Four of the strains were of the inducible type, and the remaining strains were of the multi-copy suppression type. Two of the inducible-type strains, NOT 1 and NOT40, harbored genes which had been inserted in reverse direction, suggesting that the mutant phenotypes had been derived from an excess amount of anti-sense mRNA. Domain analyses of the deduced polypeptides from the DNA fragments rescued from the transformants revealed that NOT1, NOT40 and NOT6 harbored a LisH motif, a forkhead domain, and a $Zn(II)_2Cys_6$ binuclear zinc cluster, respectively.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.662-662
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• 2005

• Proceedings of the Zoological Society Korea Conference
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• 1996.10a
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• pp.222.1-222
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• 1996

No Abstract, See Full Text