• 한국자원식물학회지
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• 제18권3호
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• pp.441-449
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• 2005

광계II(PSII)는 고등식물의 chloroplast에서 두 개의 광합성 반응중심 중의 하나이다. Chlorophyll a/b 광수확 복합체는 광계II를 위한 안테나 역할을 수행한다. 본 연구에서는 인삼의 잎조직을 제작한 cDNA library로부터 chlorophyll a/b-binding protein (Cab) 유전자를 분리하였다. 인삼 Cab유전자는 935 bp의 염기와 265개의 아미노산 잔기(pI 5.63)로 구성된 한 개의 ORF를 포함하고 있으며, 단백질의 분자량은 28.6 kDa으로 추정되었다. 인삼에서 분리한 Cab 유전자는 기존에 식물에서 보고된 유전자들과 유사성을 나타내었으며, 유사도는 $68-92\%$로 나타났다. 아미노산 서열을 비교하여 유연관계를 분석한 결과 인삼의 Cab 유전자는 비교된 P. persica (AAC34983), A.thaliana (AAD28771), G. hirsutum (CAA38025), G. max (AAL29886), V. radiata (AAF89205) 등과 동일한 그룹으로 분리되었다.

• 미생물학회지
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• 제32권3호
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• pp.215-221
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• 1994

Bacillus megaterium ATCC 14945의 penicillin G acylase 유전자의 염기배열을 결정하였다. 이 유전자에는 2,406 염기쌍으로 이루어진 하나의 open reading frame이 존재하는데, 개시코돈의 5' 위쪽에서 Shine-Dalgarno 배열과 promoter로 여겨지는 부분을 발견하였으며, 종결코돈의 3' 아래쪽에서 rho-independent한 전사종결체와 dby사한 구조를 발견하였다. 염기배열로부터 폴리펩티드의 아미노산 배열을 유추하였다. 이 폴리펩티드의 분자량은 91,983 Da이었으며, 아미노 말단 부이에 signal sequence가 존재하였다. 이 아미노산 배열을 여러 다른 penicillin G acylase의 아미노산 배열과 비교하고 분리 정제한 효소를 SDS-polyacrylamide gel 전기영동으로 분석한 결과로부터 이 효소는 92kDa의 전구체로 해독된 후 processing 과정을 거쳐 각각 25kDa과 61kDa의 ${\alpha}$-, ${\beta}$-단위체로 구성됨을 알 수 있었다.

• Journal of Photoscience
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• 제9권2호
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• pp.269-271
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• 2002

To investigate the visual and extra-ocular photoreception, we cloned the opsin genes in ayu (Plecoglossus allivelis). Amplified fragments encoding exon-4 (-5) of opsin cDNAs were cloned from the retina and brains of ayu, and sequenced. One clone was identified as rod (AYU-Rh), two as green cone (AYU-GI, -G2), one as red cone (A YU-R), two as ultraviolet cone (AYU-UVl, UV2), one as VA (AYU-VA), and one as extra-ocular rod (AYU-ExoRh) opsins. 335 amino acids sequence deduced from the full-length cDNA of AYU-Rh showed high identity with that of other fish. Southern blotting analysis indicated that ayu possess two 'rhodopsin' genes, one is visual rhodopsin and the other is non-visual extra-ocular rhodopsin. In situ hybridization showed that the mRNA of AYU-Rh was localized only in rod cells in the retina. On the other hands, AYU-ExoRh was expressed only in the pineal. We cloned two isoforms (AYU-VAM and -VAL) of VA opsin from ayu. The deduced amino acid sequences of these variants were identical to each other within the first 342 residues, but they showed divergence in the C-terminal sequence. AYU- VAL corresponded to the long isoform found in other fish, and AYU-VAM was identified as a new type of VA opsin variant. Pal-VAM is a new probably functional non-visual photoreceptive molecule in fish.

• Journal of Microbiology and Biotechnology
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• 제11권4호
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• pp.668-676
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• 2001

Pseudomonas sp. strain SY5 is a PCB-degrading bacterium [24] that includes two different enzymes (BphC1 and BphC2) encoding 2,3-dihdroxybiphenyl 1,2-dioxygenase and BphD encoding 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase. The bphC1 and bphC2 genes were found to consist of 897 based encoding 299 amino acids and 882 bases encoding 294 amino acids, respectively, whereas the bphD gene consisted of 861 bases encoding 287 amino acids. According to a homology search, a 50% and 39% similarity between the bphC1 and bphC2 genes at the nucleotide and amino acid level was shown, respectively. The bphC1 gene showed a 38% and 45% similarity at the amino acid level to Alcaligenes eutrophus A5 and Rhodococcus rhodochrous, respectively, whereas, bphC2 showed a 95% and 43% similarity, respectively. A comparison of the deduced amino acid sequence of the bphD product of Pseudomonas sp. SY5 with that of A. eutrophus A5, Pseudomons sp. KKS102, and LB400 showed a sequence identity of 92, 92, and 79%, respectively. Strain SY5 was originally isolated from municipal sewage containing recalcitrant organic compounds an found to have a high degradability of various aromatic compounds [23]. The current study found that strain SY5 had two extradiol-type dioxygenases, which did not hybridize with each other as they had a low similarity, yet a similar structure of evolutionarily conserved amino acids residues for catalytic activity between BphC1 and BphC2 was observed.

• Journal of Microbiology and Biotechnology
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• 제5권3호
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• pp.117-124
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• 1995

A gene (xynA) encoding the endo-xylanase (E.C.3.2.1.8) from Bacillus stearothermophilus was cloned in E. coli, and its complete nucleotide sequence was determined. The xynA gene consists of a 636 base pairs open reading frame coding for a protein of 212 amino acids with a deduced molecular weight of 23, 283 Da. A putative signal sequence of 27 amino acid residues shows the features comparable with the Bacillus signal sequences; namely, the signal contains a positively charged region close to the N-terminus followed by a long hydrophobic string. The coding sequence is preceded by a possible ribosome binding site with a free energy value of -16.6 kcal/mol and the transcription initiation signals are located further upstream. The translation termination codon (TAA) at the 3 end of the coding sequence is followed by two palindrome sequences, one of which is thought to act as a terminator. The xynA gene has a high GC content, especially in the wobble position of codons (64%). Comparison of the primary protein sequence with those of other xylanases shows a high homology to the xylanases belonging to family G.

• BMB Reports
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• 제41권1호
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• pp.86-91
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• 2008

The full-length cDNA of CaAbsi1 encodes a presumptive protein of 134 amino acid residues that has homology to a putative zinc finger protein in its C-terminus. The deduced amino acid sequence has 50% homology to Oryza sativa NP001049-274, the function of which is unknown. Expression of CaAbsi1 was reduced in response to inoculation of non-host pathogens. On the other hand it was induced one hour after exposure to high concentrations of NaCl or mannitol, and six hours after transfer to low temperature. Induction also occurred in response to oxidative stress, methyl viologen, hydrogen peroxide and abscisic acid. Our results suggest that CaAbsi1 plays a role in multiple responses to wounding and abiotic stresses.

• Journal of Microbiology and Biotechnology
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• 제23권6호
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• pp.872-877
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• 2013

In a previous study, we isolated and reported a second species of the Saccharophagus genus, Saccharophagus sp. strain Myt-1. In the present study, an alginate lyase gene (algMytC) from the genomic DNA of Myt-1 was cloned and characterized. The DNA sequence fragment obtained contained an open reading frame of 1,032 bp that encoded a protein of 343 amino acids with an estimated molecular mass of 37.6 kDa and a pI of 6.60. The deduced protein, AlgMytC, had the conserved amino acid sequences (RTELREM, QIH, YFKAGVYNQ) of the polysaccharide lyase family 7. A BLAST homology search indicated that AlgMytC shared an amino acid sequence identity of 95.9% with alg7A of S. degradans 2-40. The cloned and purified AlgMytC protein showed optimal activity at $40^{\circ}C$, and retained more than 90% of its total activity even after treatment at $25^{\circ}C$ for 24 h. AlgMytC was very alkaliphilic with an optimal pH of 9.0, and more than 90% of its activity was retained in the pH range 8.5-10.0. Moreover, AlgMytC was stable over a wide pH range. The activity of AlgMytC was also stable in the presence of various detergents.

• The Plant Pathology Journal
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• 제27권1호
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• pp.33-36
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• 2011

The genome of a potyvirus isolate associated with chlorotic spots and vein banding symptoms on Spathiphyllum spp., in Andhra Pradesh state, India was amplified by RT-PCR using degenerate potyvirus primers, amplicons cloned, and sequence (1.6 kb) analyzed. This virus isolate shared maximum identity of 74.8% and 80.2% at coat protein (CP) gene nucleotide (906 nucleotides) and amino acid (302 amino acids) levels, respectively with Dasheen mosaic virus (DsMV)-M13 isolate reported from China. But its 3'-UTR (258 nucleotides) had maximum identity of 62.5% with DsMV-Vietnam isolate. The deduced molecular weight of CP is 33.57 kDa and it contained DAG triplet in its N-terminal region. In CP amino acid based phylogenetic analysis, this virus isolate represented a separate branch but closer to DsMV isolates cluster. Based on the molecular criteria set for the discrimination of species and genus in the Potyviridae family, the present virus isolate was identified as a distinct virus species in the genus Potyvirus and proposed the name Spathiphyllum chlorotic vein banding virus (SCVbV).

• The Plant Pathology Journal
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• 제16권2호
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• pp.118-124
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• 2000

The coat protein (CP) gene of kyuri green mottle mosaic virus zucchini strain (KGMMV-Z) isolated from zucchini (Cucurbita pepo) in Chonfu, Korea in 1999 was sequenced by the reverse transcription and polymerase chain reaction with degenerate and generate primers originated from tobamoviruses. The degenerate primers were very effective in amplification of KGMMV-Z CP region. The KGMMV-Z CP gene consisted of 486 nucleotides and had the same nucleotide length compared with those of cucurbit-infecting tobamoviruses. KGMMV-Z CP gene shared 43.8, 44.2, and 44.4% nucleotide sequence similarity with the CP gene of cucumber green mottle mosaic virus watermelon strain (CGMMZ-W), CGMMV-KW1, and CGMMV-SH, respectively, whereas three CGMMV strains among themselves showed 98.6-99.6% nucleotide similarity. The deduced amino acids of KGMMV-Z CP gene were 161 amino acid residues with the molecular weight of 17,181 daltons. The first 24 codons of KGMMV-Z CP gene corresponded to the sequences of the N-terminal amino acid of the viral capsid protein. The amino acid sequences of KGMMV-Z CP had 45.3% similarity compared with those of three CGMMV strains. However, the amino acid sequences of CGMMV strains were identical. These results showed that two cucurbit-infecting tobamovirus members, KGMMV-Z and CGMMV were genetically distantly related.

• BMB Reports
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• 제35권2호
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• pp.213-219
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• 2002

Malonyl-CoA decarboxylase (E.C.4.1.1.9) catalyzes the conversion of malonyl-CoA to acetyl-CoA. Although the metabolic role of this enzyme has not been fully defined, it has been reported that its deficiency is associated with mild mental retardation, seizures, hypotonia, cadiomyopathy, developmental delay, vomiting, hypoglycemia, metabolic acidosis, and malonic aciduria. Here, we isolated a cDNA clone for malonyl CoA decarboxylase from a rat brain cDNA library, expressed it in E. coli, and characterized its biochemical properties. The full-length cDNA contained a single open-reading frame that encoded 491 amino acid residues with a calculated molecular weight of 54, 762 Da. Its deduced amino acid sequence revealed a 65.6% identity to that from the goose uropigial gland. The sequence of the first 38 amino acids represents a putative mitochondrial targeting sequence, and the last 3 amino acid sequences (SKL) represent peroxisomal targeting ones. The expression of malonyl CoA decarboxylase was observed over a wide range of tissues as a single transcript of 2.0 kb in size. The recombinant protein that was expressed in E. coli was used to characterize the biochemical properties, which showed a typical Michaelis-Menten substrate saturation pattern. The $K_m$ and $V_{max}$ were calculated to be $68\;{\mu}M$ and $42.6\;{\mu}mol/min/mg$, respectively.