| 金黄色葡萄球菌FnBP配体结合区基因的克隆及其原核表达(英文) |
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| 引用本文： | 尹荣兰,杨正涛,张艳晶,刘辉,刘珊,杨琦,曹永国,张乃生.金黄色葡萄球菌FnBP配体结合区基因的克隆及其原核表达(英文)[J].农业科学与技术,2008,9(6):43-46. |
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| 作者姓名： | 尹荣兰 杨正涛 张艳晶 刘辉 刘珊 杨琦 曹永国 张乃生 |
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| 作者单位： | 吉林大学畜牧兽医学院,吉林长春,130062 |
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| 基金项目： | 国家自然科学基金，教育部高等学校博士学科点专项科研基金 |
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| 摘 要： | Objective] The study aimed to clone the FnBP ligand binding gene of Staphylococcus aureus and run prokaryotic expression by constructing a prokaryotic expression vector. Method] The gene encoding FnBP ligand binding gene was amplified from S.aureus chromosomal DNA by PCR technique. After T-A cloning, plasmid pMD18- FnBP was constructed. pMD18- FnBP and pET28a(+)were digested by BamH Ⅰ and EcoR Ⅰ double enzymes, then the purified FnBP ligand binding gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-FnBP was thus constructed. The constructed plasmid pET28a-FnBP was transformed into Escherichia coli BL21(DE3) competent cells. The bacterium was induced by IPTG and the expressed products were analyzed by SDS-PAGE and Western blot. Result] The gene fragment with the length of 370 bp was amplified by PCR approach. One approximately 30 kD exogenous protein was observed in SDS-PAGE analysis. Western blot analysis indicates the protein has antigenicity of S.aureus. Conclusion] The FnBP ligand binding gene of S.aureus was successfully cloned and expressed in prokaryotic cells.
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| 关 键 词： | Staphylococcus aureus FnBP ligand binding gene Cloning Prokaryotic expression |
Cloning and Prokaryotic Expression of FnBP Ligand Binding Gene of Staphylococcus aureus |
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| YIN Rong-lan,YANG Zheng-tao,ZHANG Yan-jing,LIU Hui,LIU Shan,YANG Qi,CAO Yong-guo,ZHANG Nai-sheng.Cloning and Prokaryotic Expression of FnBP Ligand Binding Gene of Staphylococcus aureus[J].Agricultural Science ＆ Technology,2008,9(6):43-46. |
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| Authors: | YIN Rong-lan YANG Zheng-tao ZHANG Yan-jing LIU Hui LIU Shan YANG Qi CAO Yong-guo ZHANG Nai-sheng |
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| Institution: | College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062 |
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| Abstract: | Objective] The study aimed to clone the FnBP ligand binding gene of Staphylococcus aureus and run prokaryotic expression by constructing a prokaryotic expression vector. Method] The gene encoding FnBP ligand binding gene was amplified from S.aureus chromosomal DNA by PCR technique. After T-A cloning, plasmid pMD18- FnBP was constructed. pMD18- FnBP and pET28a(+)were digested by BamH Ⅰ and EcoR Ⅰ double enzymes, then the purified FnBP ligand binding gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-FnBP was thus constructed. The constructed plasmid pET28a-FnBP was transformed into Escherichia coli BL21(DE3) competent cells. The bacterium was induced by IPTG and the expressed products were analyzed by SDS-PAGE and Western blot. Result] The gene fragment with the length of 370 bp was amplified by PCR approach. One approximately 30 kD exogenous protein was observed in SDS-PAGE analysis. Western blot analysis indicates the protein has antigenicity of S.aureus. Conclusion] The FnBP ligand binding gene of S.aureus was successfully cloned and expressed in prokaryotic cells. |
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| Keywords: | Staphylococcus aureus FnBP ligand binding gene Cloning Prokaryotic expression |
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