• KSBB Journal
• /
• 제9권2호
• /
• pp.165-173
• /
• 1994

재조합 소성장호르몬을 트립신, S.aureus V8 단백질가수분해효소, CNBr, 그리고 산 가수분해법을 이용하여 단백질 일차구조 분석을 실시하였다. N-말단 분석은 30 잔기까지를 수행하였는데, 대장균 내에서 발현된 소성장호르몬은 E. coli 내 에 존재 하는 methionyl-aminopeptidase에 의해 해독개시인자로 넣어준 N-말단의 Met이 모두 제거된 형태로 나타났으며 아미노산 조성분석 결과 연역된 조성과 유사하게 나타났다. 효소와 화학물질로 절단한 소성장호르몬 조각들을 HPLC로 분리한 후 단백질 서열분석기를 이용하여 아미노산 서열을 분석하였다. 대장균에서 발현된 소성장호르몬은 191개의 아미노산으로 구성된 21,802 Da의 분자량을 갖고 있는 단백질로 나타났다. 여기에서 을 갖고 있는 단백질로 나타났다. 여기에서 얻은 아미노산 서열을 바탕으로 hydropathy plot을 한 결과 N-말단에서는 소수성이 그리고 C-말단에서는 친수성 영역이 나타났다.

• 한국수산과학회지
• /
• 제36권5호
• /
• pp.442-448
• /
• 2003

Melanin concentrating hormone (MCH) regulating color change of fish skin was identified from brain cDNA library of Olive flounder (Paralichthys olivaceus) during the analysis of Expressed Sequence Tags (ESTs). Olive flounder MCH gene consisted of 598 nucleotides encoding 150 amino acids. Olive flounder MCH protein revealed to contain signal peptide of 19 amino acid residues, pro-MCH of 131 amino acids being processed to biologically active and mature form of hormone with 25 amino acid residues at the carboxyl terminus. A comparative structural analysis revealed that Olive flounder MCH precursor had low sequence identity with other fish species and mammalian counterparts, while the amino acid sequences of mature hormone had a relatively high identity and more conserved. RT-PCR analysis revealed that olive flounder MCH precersor gene was expressed spectically only in the brain and not in other tissues.

• Journal of Microbiology and Biotechnology
• /
• 제3권3호
• /
• pp.139-145
• /
• 1993

The nucleotide sequence of the xylanase gene (xynS) from alkali-tolerant Bacillus sp. YA.14 was determined and analyzed. A 639 base pairs open reading frame for xynS gene was observed and encoded for a protein of 213 amino acids with a molecular weight of 23, 339. S1 nuclease mapping showed that the transcription initiation site of the xynS gene did not exist in the cloned DNA. Ribosome binding site sequence with the free energy of -18.8 Kcal/mol was observed 8 base pairs upstream from the initiation codon, ATG. The proposed signal sequence consisted of 28 amino acids, of which 3 were basic amino acid residues and 21 were hydrophobic amino acid residues. When the amino acid sequences of xylanases were compared, Bacillus sp. YA-14 xylanase showed 48% homology with Bacillus sp. YC-335 xylanase and 96% homology with xylanases from B. subtilis and B. circulans.

• Plant Resources
• /
• 제8권2호
• /
• pp.110-115
• /
• 2005

Ribosomal protein complex with ribosomal RNA to form the subunits of the ribosome serve essential functions in protein synthesis. A full-length cDNA (PRPS4) encoding ribosomal protein S4 has been isolated and its nucleotide sequence determined in ginseng plant (Panax ginseng). A PRPS4 cDNA is 1105 nucleotides long and has an open reading frame of 792 bp with a deduced amino acid sequence of 264 residues (pI 10.67). The deduced amino acid sequence of PRPS4 matched to the previously reported ribosomal protein S4 genes. Their degree of amino acid identity ranged from 68 to $92\%$. Phylogenetic analysis based on the amino acid residues showed that the PRPS4 grouped with ribosomal protein S4 of S. tuberosum (CAA54095).

• 미생물학회지
• /
• 제41권1호
• /
• pp.13-17
• /
• 2005

본 연구에서는 이 전 연구에서 단일 탄소원과 질소원 및 에너지원으로 aniline을 이용하는 Delftia sp. JK-2에서 분리, 정제된 바 있는 C2,3O의 N-말단 아미노산과 DNA 서열을 분석하였다. Aniline에서 배양한 Delftia sp. JK-2에서 분리된 약 35kDa의 C2,3O의 N-말단 아미노산 서열을 분석 한 결과 $^1MGVMRIGHASLKVMDMDAAVRHYENV^{26}$로 Pseudomonas sp. AW-2와 Comamonas sp. JS765의 C2,3O와 일치하는 것으로 나타났다. 위에서 확인된 아미노산 서열을 바탕으로 제작된 primer와 JK-2의 total genomic DNA를 기질로 사용하여 PCR을 수행한 결과 약 950 bp의 유전자 증폭산물을 획득하였다. 이 증폭산물 중 정확히 확인된 890 bp의 염기서열을 분석한 결과 Delftia JK-2의 C2,3O유전자 염기서열은 Pseudomonas su. AW-2의 C2,3O와 일치하였으며 Comamonas sp. Js765의 C2,3O와 $97\%$의 높은 상동성을 나타내었다.

• Journal of Microbiology and Biotechnology
• /
• 제8권5호
• /
• pp.531-535
• /
• 1998

The gene encoding alginate lyase was isolated from a library constructed with the vector, pUC19, and expressed in Escherichia coli. The nucleotide sequence of the cloned alginate lyase gene (ALY) from Pseudomonas sp. W7 was determined. The nucleotide sequence revealed a 1,035 bp open reading frame (ORF), encoding 345 amino acid residues with a calculated molecular mass of 37,478 Da. The N-terminal amino acid sequences (15 residues) of purified alginate lyase corresponded to that of the deduced amino acid sequence.

• Animal cells and systems
• /
• 제12권3호
• /
• pp.149-155
• /
• 2008

We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

• Journal of Microbiology and Biotechnology
• /
• 제3권4호
• /
• pp.224-231
• /
• 1993

The nucleotide sequence of the xylanase gene (xynY) from alkalophilic Bacillus sp. YC-335 was determined and analyzed. An open reading frame of 1, 062 base pairs for xynY gene was observed and encoded for a protein of 354 amino acids with a molecular weight of 38, 915. S1 nuclease mapping showed that the transcription initiation sites of the xynY gene were different in Bacillus sp. YC-335 and Escherichia coli HB101 (pYS55). S1 mapping also showed that -10 region of the xynY gene recognized by RNA polymerases of E. coli and Bacillus sp. YC-335 were TACAGT and TATGAT , respectively. A ribosome binding site sequence with the free energy of -17.0 Kcal/mol was observed 9 base pairs upstream from the unusual initiation codon, TTG. The proposed signal sequence consisted of 27 amino acids, 2 of which were basic amino acid residues and 21 were hydrophobic amino acid residues. When the amino acid sequences of xylanases were compared, Bacillus sp. YC-335 xylanase showed more than 50% homology with xylanases from B. pumilus, B. subtilis, and B. circulans.

• 약학회지
• /
• 제34권1호
• /
• pp.11-14
• /
• 1990

$NH_2-terminal$ amino acid sequence analysis of monoclonal antibody is very important to identify gene family and diversities of antigen-antibody recognition. When we used the PVDF (Polyvinylidene difluoride) membrane blotting method, we could easily analyze $NH_2-terminal$ sequence of monoclonal-antibody which specifically binds to phosphatidylinositol 4,5-biphosphate. PVDF membrane is an ideal solid-phase support for sequence analysis, especially when used with electroblotting method. This method is superior to continual method and will be applied to the sequence analysis of picomole quantities of proteins by gel electrophoresis.

• The Plant Pathology Journal
• /
• 제16권4호
• /
• pp.231-235
• /
• 2000

The 3,000 nucleotides of 3'-terminal region of the genomic RNA of a new isolate of carlavirus from a Korean native lily (Lilum lancitoium) was cloned and its nucleotide sequences were determined. The coat protein (CP) gene of the virus showed 72.0% to 72.8% nucleotide sequence identities and 86.9% to 88.0% amino acid sequence identities with those of the four strains (two Korean, one Dutch, and one Japanese isolates) of lily symptomless virus (LSV). Interestingly, different amino acid sequences between the new isolate and LSV strains were located at the N-terminal region of the CP. Pairwise amino acid sequence comparison of the CP gene revealed sequence identities of 22.0% to 71.1% between the virus and other 9 carlavirus species. The 25 kDa and 12 kDa proteins genes of the virus share 30.7% to 76.3% and 31.1% to 85.8% amino acid sequence identities, respectively, with those of 8 other carlaviruses. The 16 kDa protein gene of the virus shares 16.7% to 72.9% amino acid sequence identities with that of 9 other carlaviruses. These data indicate that the virus, designated as lily latent virus (LiLV), is a distinct of the Carlavirus genus and distinguished from the known strains of LSV.