• Journal of Microbiology and Biotechnology
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• v.24 no.10
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• pp.1389-1396
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• 2014

The entire nucleotide sequence of the TKL1 gene encoding transketolase (TKL) in an erythritol-producing yeast of Candida magnoliae was determined by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed an open reading frame of C. magnoliae TKL1 (CmTKL1) that spans 2,088 bp and encodes 696 amino acids, sharing 61.7% amino acid identity to Kluyveromyces lactis TKL. Functional analysis showed that CmTKL1 complemented a Saccharomyces cerevisiae tkl1 tkl2 double mutant for growth in the absence of aromatic amino acids and restored transketolase activity in this mutant. An enzyme activity assay and RT-PCR revealed that the expression of CmTKL1 is induced by fructose, $H_2O_2$, and KCl. The GenBank accession number for C. magnoliae TKL1 is KF751756.

• Journal of Life Science
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• v.9 no.2
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• pp.9-14
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• 1999

Pseudomonas iodinum IFO 3558 adenosine deaminase(ADA) gene was cloned by the polymerase chain reaction and deduced the amino acid sequence of the enzyme. DNA sequence homology of Pseudomonas iodinum IFO 3558 ADA gene was compared to those of E. coli, human and mouse ADA genes. Unambiguous sequence from both strands of pM21 was obtained for the region believed to encode ADA. The sequence included a 804-nucleotide open reading frame, bounded on one end by sense primer and on the other end by two antisense primer. This open reading frame encodes a protein of 268 amino acids having a molecular weight of 29,448. The deduced amino acid sequence shows considerable similarity to those of E. coli, mouse and human ADA. Pseudomonas iodinum IFO 3558 nucleotide sequence shows 98.5% homology with that of the E. coli ADA sequence and 51.7% homology with that of the mouse ADA sequence and 52.5% homology with that of the human ADA sequence. The ADA protein sequence of Pseudomonas iodinum IFO 3558 shows 96.9% homology with that of the E. coli and 40.7% homology with that of the mouse and 41.8% homology with that of the human. The distance between two of the conserved elements, TVHAGE and SL(1)NTDDP has veen exactly conserved at 76 amino acids for all four ADAs. Two of the four conserved sequence elements shared among the four ADAs are also present in the yeast, rat, human (M), and Human(L) AMP deaminase. The SLSTDDP sequence differs only in the conservative substitution of a serine for an asparagine. A conserved cysteine with conserved spacing between these two regions is also found. Thus, sequence analysis of four ADAs and four AMP deaminases revealed the presence of a highly conserved sequence motif, SLN(S)TDDP, a conserved dipeptide, HA, and a conserved cysteine residue.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.11
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• pp.1673-1679
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• 2008

In order to study the biological function of gapdh gene in yak, and prove whether the gapdh gene was a useful intra-reference gene that can be given an important role in molecular biology research of yak, the cDNA sequence encoding glyceraldehyde-3-phosphate dehydrogenase from yak was cloned by the RT-PCR method using gene specific PCR primers. The sequence results indicated that the cloned cDNA fragment (1,008 bp) contained a 1,002 bp open reading frame, encoding 333 amino acids (AAs) with a molecular mass of 35.753 kDa. The deduced amino acids sequence showed a high level of sequence identity to Bos Taurus (99.70%), Xenopus laevis (94.29%), Homo sapiens (97.01%), Mus musculus (97.90%) and Sus scrofa (98.20%). The expression of yak's gapdh gene in heart, spleen, kidney and brain tissues was also detected; the results showed that the gapdh gene was expressed in all these tissues. Further analysis of yak GAPDH amino acid sequence implied that it contained a complete glyceraldehyde-3-phosphate dehydrogenase active site (ASCTTNCL) which ranged from 148 to 155 amino acid residues. It also contained two conserved domains, a NAD binding domain in its N-terminal and a complete catalytic domain of sugar transport in its C-terminal. The phylogenetic analysis showed that yak and Bos taurus were the closest species. The prediction of secondary structures indicated that GAPDH of yak had a similar secondary structure to other isolated GAPDH. The results of this study suggested that the gapdh gene of yak was similar to other species and could be used as the intra-reference to analyze the expression of other genes in yak.

• Journal of fish pathology
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• v.19 no.3
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• pp.227-233
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• 2006

Olive flounder immunoreceptor DAP10 homologue cDNA was cloned from a peripheral blood lymphocytes (PBLs) cDNA library. The length of the olive flounder DAP10 cDNA is 473bp and it contains an open reading frame of 234bp. The predicted polypeptide sequence is 78 amino acids, consisting of a 22-amino acid leader, an 11-amino acid extracellular domain, a 21-amino acid transmembrane segment, and a 24-amino acid cytoplasmic domain. The amino acid sequence of olive flounder DAP10 has 56%, 50%, 32%, 31%, and 31% sequence identity with zebrafish DAP10, catfish DAP10, cattle DAP10, rat DAP10 and Monkey DAP10, respectively. Olive flounder DAP10 has a conserved aspartic acid in the transmembrane domain and a phophatidylinositol-3 kinase-binding site (YxxM/V) in the cytoplasmic region. Genomic organization reveals that olive flounder DAP10 comprises five exons and four introns. A phylogenetic analysis based on the deduced amino acid sequence grouped the olive flounder DAP10 with other species DAP10. In RT-PCR analysis, DAP10 transcripts were detected predominantly in PBLs, kidney, spleen and intestine.

• Fisheries and Aquatic Sciences
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• v.4 no.4
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• pp.192-196
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• 2001

Proopiomelanocortin (POMC) plays an essential role in the disease resistance system and is the precursor protein of biologically active peptides such as adrenocorticotropin (ACTH), $\alpha-melanocyte-stimulating$ hormone $(\alpha- MSH)$, $(\beta-melanocyte-stimulation hormone\;(\beta- MSH)$ and $\beta-endorphin$. We have isolated and sequenced two different forms of POMC cDNA, POMC-I and POMC-II, from a pituitary cDNA library of flounder. POMC-I cDNA consisted of 956 bp corresponding to deduced amino acids of 216 residues and POMC-II cDNA was 982 bp in length corresponding to 194 amino acids, respectively. The results of deduced amino acids analysis of the clones showed high sequence homology with previously reported POMCs amino acid sequences from various species. The homology between flounder POMC-I and -II is$57\%$ identity. We also constructed a phylogenetic tree based on POMC amino acid sequences.

• Korean Journal Plant Pathology
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• v.14 no.3
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• pp.240-246
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• 1998

A cDNA encoding desiccation-related protein was isolated from a flower bud cDNA library of Chinese cabbage (C-DH) and its nucleotide sequence was characterized. It contains 679 bp nucleotides with 501 bp open reading frame. The amino acid sequence of the putative protein showed the highest amino acid sequence homology (79 % identity) to dehydrin protein in Gossypium hirsutum. Also, the C-DH shares 48-52% amino acid sequence identity with the other typical dehydrin proteins in plant cells. When the amino acid sequence of their proteins were aligned, several peptide motifs were well conserved, of which function has to be solved. Particularly the C-DH contains 15 additional amino acids at its N-terminus. Genomic Southern blot analysis using the coding region of C-DH showed that the C-DH consists of a single copy gene in Chinese cabbage genome. The C-DH mRNA, whose transcript size is 0.7 kb, was expressed with a tissue-specific manner. It was highly expressed in seed, flower buds and low expression as detected in root, stem or leaf tissues of Chinese cabbage. And the transcript level of C-DH was significantly induced by the treatment of plant hormone, abscisic acid and water-deficit conditions.

• Korean Journal Plant Pathology
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• v.14 no.4
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• pp.314-318
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• 1998

Using the reverse transcription polymerase chain reaction (RT-PCR), a rapid and sensitive assay method for the detection and identification of barley yellow mosaic virus (BaYMV) and barley mild mosaic virus (BaMMV) was adapted. Two units of primers from each virus were selected and used for the determination of two different viruses. PCR fragments of BaYMV (ca. 0.9kb) and BaMMV (ca. 0.8kb) were obtained from the designed method for the assay of BaYMV and BaMMV coat protein. PT-PCR fragments were cloned using vector pT7 Blue and the sequences of the selected clones were analyzed. coat protein of BaYMV and that of BaMMV consisted of 297 amino acids (891 nucleotides) and 251 amino acids (753 nucleotides), respectively. The snalysis of coat protein genes from these two viruses showed that 45.6% of nucleotides sequence ad 34.9% of amino acid in BaYMV were homologous to those in BaMMV.

• Journal of Microbiology and Biotechnology
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• v.2 no.1
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• pp.56-65
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• 1992

The complete nucleotide sequence of the Bacillus circulans F-2 RSDA gene, coding for raw starch digesting a-amylase (RSDA), has been determined. The RSDA structure gene consists of an open reading frame of 2508 bp. Six bp upstream of the translational start codon of the RSDA is a typical gram-positive Shine-Dalgarno sequence and the RSDA encodes a preprotein of 836 amino acids with an Mr of 96, 727. The gene was expressed from its own regulatory region in E. coli and two putative consensus promoter sequences were identified upstream of a ribosome binding site and an ATG start codon. Confirmation of the nucleotide sequence was obtained and the signal peptide cleavage site was identified by comparing the predicted amino acid sequence with that derived by N-terminal analysis of the purified RSDA. The deduced N-terminal region of the RSDA conforms to the general pattern for the signal peptides of secreted prokaryotic proteins. The complete amino acid sequence was deduced and homology with other enzymes was compared. The results suggested that the Thr-Ser-rich hinge region and the non-catalytic domain are necessary for efficient adsorption onto raw substrates, and the catalytic domain (60 kDa) is necessary for the hydrolysis of substrates, as suggested in previous studies (8, 9).

• Plant Resources
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• v.2 no.2
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• pp.69-74
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• 1999

A rapid and sensitive assay for specific detection and identification of barley yellow mosaic virus(BaYMV) was set up using the reverse transcriptase polymerase chain reaction(RT-PCR). A couple of primers was select to discriminate the viruses. PCR fragments of BaYMV(ca.0.9 kb) were obtained by using the method designed for BaYMV capsid protein. RT-PCR fragments were cloned with vector pT7 Blue and the resulting clones were sequenced. Capsid protein of BaYMV consisted of 297 amino acids and 891 nucleotides. The capsid protein sequence of BaYMV showed that 98% of nucleotides and 99% of amino acids homology.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.194-200
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• 1989

The nucleotide sequence of inducible chloramphenicol acetyl-transferase(CAT) gene isolated from a small plasmid pSBK203 of Staphylococcus aureus was determined. The base sequence shows that structural gene of pSBK203-CAT encodes a protein of 213 amino acids and has a leader region which encodes a short polypeptide of 9 amino-acids in its upstream. vertical bar /sup 35/S vertical bar-Methionine labelled CAT gene product in minicell showed almost same mobility with pC194-CAT of which molecular weight is 24Kdal on polyacrylamide gel electrophoresis. Predicted amino acid sequence of pSBK203-CAT has revealed a high degree of homology with the CATs of pC194 and pC221 than those of cat-86, Tn9 and proteus mirabilis PM13.