中国人兽共患病学报2011，Vol.27Issue(4)：287-290,4.
猪附红细胞体ORF4基因片段的人工合成及基因表达
Cloning and expressing of synthesis fragment of Mycoplasma suis ORF4 gene
摘要
Abstract
In this research, the ORF4 gene of the Mycoplasma suis was synthesized and put into E.coli for expression.Base on the gene sequence of the ORF4 gene of the AJ504999 which registered in the GenBank, according to the Graphical Codon Usage Analyser, three pairs of oligonucleotides were synthesized by using E.coli biased codons and changing transl_table.The fragment was amplified by PCR-based accurate synthesis (PAS).Then it was inserted into expression vector PET28a after sequenced.The correctly recombinant plasmid PET28a/ORF4 was transformed into E.coli BL21.The recombinant was induced under 30℃ and 1 mmol/L concentration of IPTG.About 11kD a protein was appeared obviously by SDS-PAGE.The bioinformatics software analytic results showed that there were nineteen binding site residues, and a most likely cleavage site between position 21 and 22 AA in the ORF4 protein model.It was intracellular protein and had not glycosylation site.Consequently, ORF4 gene could be expressed in E.coli, suggesting that it has important roles in the formation of the protein and the biological function in M.suis.关键词
猪附红细胞体/ORF4/全基因合成/表达分类
生物科学引用本文复制引用
周栋,陈甜甜,刘建柱,成子强,张利,刘海涛,张璐,刘永夏,柴同杰..猪附红细胞体ORF4基因片段的人工合成及基因表达[J].中国人兽共患病学报,2011,27(4):287-290,4.基金项目
中国博士后基金特别资助(200801417),山东省博士后创新项目专项基金资助(200801001),山东省博士后基金资助(76247),山东省中青年科学家奖励基金(BS2009NY007) （200801417）
