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Cloning, Sequence Analysis And Prokaryotic Expression Of The Ninth Part Of ATP Synthase A Subunit Gene Atp9Unit In Trametes Gallica

Posted on:2013-05-19Degree:MasterType:Thesis
Country:ChinaCandidate:W J SiFull Text:PDF
GTID:2230330377450909Subject:Biochemistry and Molecular Biology
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As a white rot fungus, Trametes gallica is able to produce and secrete ligninolytic enzymes and thus degrade much lignin in nature. There are a very broad application prospects in the field of environmental protection and resource recycling. ATP synthase is a key enzyme of energy metabolism, which was involved in a variety of oxidative phosphorylation and photophosphorylation in vivo. The atp9gene encoding the ninth part of ATP synthase A subunit is an important component of ATP synthase. It is closely related with respiration and photosynthesis. For its highly conserved sequence in evolution, one pair of primers were designed and synthesized according to the known gene sequences from closely related fungi. The first strand of Trametes gallica cDNA was amplified by reverse transcription and ten the complete atp9gene sequences were obtained by PCR. After that, the product of PCR was ligated to the prokaryotic expression vector pET32a (+) and transformed into E. coli for the expression of fusion protein. The sequenced and bioinformatics analysis showed that:the complete length of atp9gene is222bp, and the peptide it encoded have73amino acids with a molecular mass of7.35kD as predicted. Homology comparison and phylogenetic analysis indicated that the amino acid sequence of Trametes gallica atp9gene possessed the highest homology with Crinipellis perniciosa and Trametes cingulata. This research laid the foundation for probing the function and metabolic pathways of atpA gene in the future.
Keywords/Search Tags:atp9gene, prokaryotic expression, sequence analysis, Trametesgallica
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