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| 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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| Yang,Yanhua; Zhu,Keming; Xia,Hengchuan; Chen,Liang; Chen,Keping. |
| Indica and japonica are two main subspecies of Asian cultivated rice (Oryza sativa L.) that differ clearly in morphological and agronomic traits, in physiological and biochemical characteristics and in their genomic structure. However, the proteins and genes responsible for these differences remain poorly characterized. In this study, proteomic tools, including two-dimensional electrophoresis and mass spectrometry, were used to globally identify proteins that differed between two sequenced rice varieties (93-11 and Nipponbare). In all, 47 proteins that differed significantly between 93-11 and Nipponbare were identified using mass spectrometry and database searches. Interestingly, seven proteins were expressed only in Nipponbare and one protein was... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Indica and japonica rice; Molecular marker; Proteomics; Quantitative real-time PCR; Unique proteins. |
| Ano: 2014 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572014000500008 |
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| 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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| Yang,Yanhua; Dai,Li; Xia,Hengchuan; Zhu,Keming; Liu,Haijun; Chen,Keping. |
| Seeds are the most important plant storage organ and play a central role in the life cycle of plants. Since little is known about the protein composition of rice (Oryza sativa) seeds, in this work we used proteomic methods to obtain a reference map of rice seed proteins and identify important molecules. Overall, 480 reproducible protein spots were detected by two-dimensional electrophoresis on pH 4-7 gels and 302 proteins were identified by MALDI-TOF MS and database searches. Together, these proteins represented 252 gene products and were classified into 12 functional categories, most of which were involved in metabolic pathways. Database searches combined with hydropathy plots and gene ontology analysis showed that most rice seed proteins were hydrophilic... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Mass spectrometry; Proteomic analysis; Rice seed; Two-dimensional electrophoresis. |
| Ano: 2013 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572013000100012 |
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| 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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