• Proceedings of the Korean Environmental Sciences Society Conference
• /
• 2002.05b
• /
• pp.445-447
• /
• 2002

본 연구는 환경친화적인 biosurfactant를 생산하는 Pseudomonas aeruginosa EMS1 and KH7를 rhamnolipid의 rhlR, rhlA, rhlB를 기초로한 primer를 이용하여 752bp, 802pb, 1280bp pcr을 수행하였으며 $pGEM^{(R)}$ / - T Easy Vector gene cloning 하여 Pseudomonas aeruginosa EMS1 and KH7의 Partial Sequencing를 서로 비교하였다. 이들 실험을 통하여 Pseudomonas aeruginosa의 유전적 구조 및 특성을 비교하여 유전적 조작을 위한 기초적인 자료가 되도록 한다.

• Journal of Crop Science and Biotechnology
• /
• v.10 no.4
• /
• pp.237-242
• /
• 2007

Genome duplication(i.e. polyploidy) is a common phenomenon in the evolution of plants. The objective of this study was to achieve a comprehensive understanding of genome duplication for SNP discovery by Thymine/Adenine(TA) cloning for confirmation. Primer pairs were designed from 793 EST contigs expressed in the roots of a supernodulating soybean mutant and screened between 'Pureunkong' and 'Jinpumkong 2' by direct sequencing. Almost 27% of the primer sets were failed to obtain sequence data due to multiple bands on agarose gel or poor quality sequence data from a single band. TA cloning was able to identify duplicate genes and the paralogous sequences were coincident with the nonspecific peaks in direct sequencing. Our study confirmed that heterogeneous products by the co-amplification of a gene family member were the main cause of obtaining multiple bands or poor quality sequence data in direct sequencing. Counts of amplified bands on agarose gel and peaks of sequencing trace suggested that almost 27% of nonrepetitive soybean sequences were present in as many as four copies with an average of 2.33 duplications per segment. Copy numbers would be underestimated because of the presence of long intron between primer binding sites or mutation on priming site. Also, the copy numbers were not accurately estimated due to deletion or tandem duplication in the entire soybean genome.

• Journal of Ginseng Research
• /
• v.33 no.1
• /
• pp.55-58
• /
• 2009

Generally, the direct sequencing through PCR is faster, easier, cheaper, and more practical than clone sequencing. Frequently, standard PCR amplification is usually interpreted by mispriming internal or external regions of the target template. Normally, DNA fragments were eluted from the gel using Gel extraction kit and subjected to direct sequencing or cloning sequencing. Cloning sequencing has often troublesome and needs more time to analyze for many samples. Since touchdown (TD) PCR can generate sufficient and highly specific amplification, it reduces unwanted amplicon generation. Accordingly, TD PCR is a good method for direct sequencing due to amplifying wanted fragment. In plants the 5S-rRNA gene is separated by simple spacers. The 5S-rRNA gene sequence is very well-conserved between plant species while the spacer is species-specific. Therefore, the sequence has been used for phylogenetic studies and species identification. But frequent occurrences of spurious bands caused by complex genomes are encountered in the product spectrum of standard PCR amplification. In conclusion, the TD PCR method can be applied easily to amplify main 5S-rRNA and direct sequencing of panax ginseng cultivars.

• Korean Journal of Microbiology
• /
• v.40 no.1
• /
• pp.12-16
• /
• 2004

Acid proteinase has been discovered in Aspergillus niger (acid protease A) and Cryphonectria parasitica (acid proteinase EapC) and it plays major roles in cheese formation from milk. In this study, a partial gene encoding acid proteinase in Penicillium oxalicum HCLF-34 was cloned by using PCR with degenerate primers corresponding to highly conserved regions of the acid proteinase. The partial acid proteinase gene in P. oxalicum HCLF-34 contains an open reading frame of 438 base pairs and encodes an acid proteinase protein of 146 amino aicds. The predicted amino acid sequences showed 71 % homology with acid protease A and 67% homology with EapC.

• Proceedings of the KSAR Conference
• /
• 2002.06a
• /
• pp.90-90
• /
• 2002

In this study, we report the cloning of the porcine UPII genomic DNA, which contains a putative full-length open reading frame encoding the UPII protein. A comparison of the porcine UPII gene coding sequence with the previously published mouse UPII sequence demonstrates that only the exon sequences are partially conserved. Northern and immunohistochemical analyses show that the porcine UPII gene is expressed only in the urothelium and that the protein specifically localizes to urothelial superficial cells. (omitted)

• Ocean and Polar Research
• /
• v.27 no.2
• /
• pp.205-214
• /
• 2005

From ancient Antarctic glacier ice, we extracted total genomic DNA that was suitable for prokaryotic 16S rDNA gene cloning and sequencing, and bacterial artificial chromosome (BAC) library and end-sequencing. The ice samples were from the Dry Valley region. Age dating by $^{40}Ar/^{39}Ar$ analysis on the volcanic ashes deposited in situ indicated the ice samples are minimum 100,000-300,000 yr (sample DLE) and 8 million years (sample EME) old. Further assay proved the ice survived freeze-thaw cycles or other re-working processes. EME, which was from a small lobe of the basal Taylor glacier, is the oldest known ice on Earth. Microorganisms, preserved frozen in glacier ice and isolated from the rest of the world over a geological time scale, can provide valuable data or insight for the diversity, distribution, survival strategy, and evolutionary relationships to the extant relatives. From the 16S gene cloning study, we detected no PCR amplicons with Archaea-specific primers, however we found many phylotypes belonging to Bacteria divisions, such as Actinobacteria, Acidobacteria, Proteobacteria $({\alpha},\;{\beta},\;and\;{\gamma})$, Firmicutes, and Cytophaga-Flavobacterium-Bacteroid$. BAC cloning and sequencing revealed protein codings highly identical to phenylacetic acid degradation protein paaA, chromosome segregation ATPases, or cold shock protein B of present day bacteria. Throughput sequencing of the BAC clones is underway. Viable and culturable cells were recovered from the DLE sample, and characterized by their 16S rDNA sequences. Further investigation on the survivorship and functional genes from the past should help unveil the evolution of life on Earth, or elsewhere, if any.

• Proceedings of the Zoological Society Korea Conference
• /
• 1995.10b
• /
• pp.322-322
• /
• 1995

• Proceedings of the Zoological Society Korea Conference
• /
• 1995.10b
• /
• pp.323.1-323
• /
• 1995

• Proceedings of the Zoological Society Korea Conference
• /
• 2000.10a
• /
• pp.284.2-284
• /
• 2000

No Abstract, See Full Text

• Proceedings of the Zoological Society Korea Conference
• /
• 1997.10b
• /
• pp.287.1-287
• /
• 1997

No Abstract, See Full Text