• 농약과학회지
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• 제10권1호
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• pp.56-65
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• 2006

작물 근권 토양으로부터 분리한 5,000여 길항 균주로부터 Erwinia 및 Pseudomonas등의 세균과 Trichoderma, Colletotrichum 등 곰팡이의 성장을 동시에 억제하는 Bacillus sp. N 32 균주를 선발 동정 하였다. 특히 Bacillus sp. N32 균주는 고추 탄저병균인 Colletotrichum gloeosporioides에 대하여 열에 저항성이 있는 단백질과 열에 민감한 단백질의 2종류의 항균 단백질을 동시에 생산함을 단백질 침전과 활성 검정을 통하여 확인하였다. 이 항균 단백질들을 FPLC를 이용한 gel filtration chromatography방법으로 분리한 후 SDS-PAGE와 bioautography로 항균력을 확인하였다. 또한 이 항균 단백질의 유전자들을 선발하기 위하여 기존의 알려진 그람양성 세균의 대표적인 열 저항성 항균 펩타이드 생합성 유전자 서열을 primer로 이용한 PCR 방법으로 fengycin의 생합성 유전자 단편을 분리하고 이 PCR 산물을 이용하여 Bacillus sp. N32 균주의 cosmid library로부터 fengycin의 생합성 유전자 cluster중 일부를 분리하여 염기서열을 분석하였다. 또한 열에 민감한 항균 단백질 생산 유전자는 이 항균 단백질을 SDS-PAGE 및 electroblotting으로 분리한 뒤 N-terminal 부위의 15개의 아미노산 서열을 분석하고 이를 DNA 염기배열로 치환한 다음 probe로 이용하여 ${\lambda}-ZAP$ library로부터 항균 단백질 생산 유전자가 포함된 다수의 clone을 선발하였다.

• Journal of Plant Biotechnology
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• 제3권3호
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• pp.131-134
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• 2001

In $\lambda$-Zap II vector system, a cDNA library was constructed for the developing secondary xylem mRNA from hybrid poplar, Populus nigra x maximowiczii. A cDNA clone of 1.5 kb in size, pOMTB1.4 encoding a lignin-bispecific O-methyltransferase was screened by plaque hybridization using a probe of 540 bp cDNA amplified by polymerase chain reaction from the cDNA library and identified by nucleotide sequencing. Its nucleotide sequence contains one open reading frame of 366 amino acids. The deduced amino acid sequence in comparison with that of Populus tremuloides showed the differences of 9 amino acids and revealed 85-99% homology among alfalfa, poplar and aspen.

• 한국수산과학회지
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• 제30권6호
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• pp.984-991
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• 1997

New tumor suppressor gene, snm23, homologous to human nm23/NDP kinase (human nucleoside diphosphate kinase) gene whose product has a tumor metastasis inhibitory activity, was first cloned from Korean tiger shark (Scyliorhinus forazame) skin cDNA library constructed by using a $\lambda$ ZAP-II cDNA synthesis kit. About $1\times10^5$ plaques were screened and several positive plaques were isolated and confirmed by second screening. The phagemid containing a positive clone from the Uni-Zap XR vector was excised in vivo and the gene containing the tumor metastasis suppressor protein was named as snm23. Cloned gene, snm23, was sequenced with ABI-PRISM 310 Genetic Analyzer. The nucleotide and deduced amino acid sequences of snm23 have shown an open reading frame consisting of 450 base pairs that correspond to a protein of 150 amino acid residues, with a calculated molecular mass of 16.8 kDa. Sequence comparison of snm23 with human nm23/NDP kinase was performed by using Blast protein data base of National Center for Biotechnology Information. In order to determine tissue specificity, reverse transcription-polymerase chain reaction (RT-PCR) was used. Good expression level of snm23/NDP kinase was detected at the tissues from skin, cartilage, and liver of Korean tiger shark.

• Parasites, Hosts and Diseases
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• 제49권1호
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• pp.79-83
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• 2011

Trichomoniasis is a sexually transmitted disease due to infection with Trichomonas vaginalis, and it can cause serious consequences for women's health. To study the virulence factors of this pathogen, T. vaginalis surface proteins were investigated using polyclonal antibodies specific to the membrane fractions of T. vaginalis. The T. vaginalis expression library was constructed by cloning the cDNA derived from mRNA of T. vaginalis into a phage ${\lambda}$ Uni-ZAP XR vector, and then used for immunoscreening with the anti-membrane proteins of T. vaginalis antibodies. The immunoreactive proteins identified included adhesion protein AP65-1, ${\alpha$-actinin, kinesin-associated protein, teneurin, and 2 independent hypothetical proteins. Immunofluorescence assays showed that AP65-1, one of the identified immunogenic clones, is prevalent in the whole body of T. vaginalis. This study led us to identify T. vaginalis proteins which may stimulate immune responses by human cells.