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Cloning, expression and characterization of alcohol dehydrogenases in the silkworm Bombyx mori Genet. Mol. Biol.
Wang,Nan; Shi,Haifeng; Yao,Qin; Zhou,Yang; Kang,Lequn; Chen,Huiqin; Chen,Keping.
Alcohol dehydrogenases (ADH) are a class of enzymes that catalyze the reversible oxidation of alcohols to corresponding aldehydes or ketones, by using either nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP), as coenzymes. In this study, a short-chain ADH gene was identified in Bombyx mori by 5'-RACE PCR. This is the first time the coding region of BmADH has been cloned, expressed, purified and then characterized. The cDNA fragment encoding the BmADH protein was amplified from a pool of silkworm cDNAs by PCR, and then cloned into E. coli expression vector pET-30a(+). The recombinant His-tagged BmADH protein was expressed in E. coli BL21 (DE3), and then purified by metal chelating affinity chromatography. The...
Tipo: Info:eu-repo/semantics/article Palavras-chave: 5'-RACE PCRADH; Enzymatic activity; Recombinant protein.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000200013
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Molecular cloning and expression of key gene encoding hypothetical DNA polymerase from B. mori parvo-like virus Genet. Mol. Biol.
Zhang,Junhong; Li,Guohui; Chen,Huiqing; Li,Xiaogang; Lv,Meng; Chen,Keping; Yao,Qin.
BmPLV-Z is the abbreviation for Bombyx mori parvo-like virus (China isolate). This is a novel virus with two single-stranded linear DNA molecules, viz., VD1 (6543 bp) and VD2 (6022 bp), which are encapsidated respectively into separate virions. Analysis of the deduced amino acid sequence of VD1-ORF4 indicated the existence of a putative DNA-polymerase with exonuclease activity, possibly involved in the replication of BmPLV-Z. In the present study, a recombinant baculovirus was constructed to express the full length of the protein encoded by the VD1-ORF4 gene (3318 bp). In addition, a 2163-bp fragment amplified from the very same gene was cloned into prokaryotic expression vector pET-30a and expressed in E.coli Rosetta 2 (DE3) pLysS. The expressed fusion...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bombyx mori parvo-like virus; DNA polymerase; Recombinant protein; Sf-9.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000400021
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Proteomic profiling of liver from Elaphe taeniura, a common snake in eastern and southeastern Asia Genet. Mol. Biol.
Chen,Liang; Xia,Hengchuan; Wang,Yiting; Chen,Keping; Qin,Lvgao; Wang,Bin; Yao,Qin; Li,Jun; He,Yuanqing; Zhao,Ermi.
Snake liver has been implicated in the adaptation of snakes to a variety of habitats. However, to date, there has been no systematic analysis of snake liver proteins. In this study, we undertook a proteomic analysis of liver from the colubrid snake Elaphe taeniura using a combination of two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flightmass spectrometry (MALDI-TOF MS). We also constructed a local protein sequence database based on transcriptome sequencing to facilitate protein identification. Of the 268 protein spots revealed by 2-DE 109 gave positive MS signals, 84 of which were identified by searching the NCBInr, Swiss-Prot and local databases. The other 25 protein spots could not be identified, possibly...
Tipo: Info:eu-repo/semantics/article Palavras-chave: 2D electrophoresis; Elaphe taeniura; MALDI-TOF mass spectrometry; Protein.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572013000300020
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