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| 康氏木霉纤维素酶CBHI基因克隆及在大肠杆菌中的表达 | |
| 引用本文： | 黄时海,康超,黄飞,曹喜秀,何鑫平,汪晟,吴孔阳,曹普美,梁智群,李湘萍. 康氏木霉纤维素酶CBHI基因克隆及在大肠杆菌中的表达[J]. 中国酿造, 2011, 0(2) |
| 作者姓名： | 黄时海 康超 黄飞 曹喜秀 何鑫平 汪晟 吴孔阳 曹普美 梁智群 李湘萍 |
| 作者单位： | 1. 广西大学生命科学与技术学院,广西,南宁,530005 2. 南宁新技术创业者中心,广西,南宁,530007 3. 广西大学动物繁殖研究所,广西,南宁,530005 |
| 基金项目： | 广西区教委人才项目，国家大学生创新基金，广西大学实验教改项目 |
| 摘 要： | 将康氏木霉(Trichoderma koningii)总RNA反转录成cDNA第一链,并以之为模板进行RT-PCR,合成约1.5kb的纤维二糖水解酶Ⅰ(cbh I)基因.cbhI基因经测序确认后克降到表达载体pET-30a(+)上,PCR和双酶切鉴定筛选阳性重组子;将阳性质粒转化大肠杆菌BL21(DE3)plysS,并用0.4mmol/L的IPTG诱导表达重组蛋白.实验结果:cbhI基因在BL21(DE3)plysS中胞内融合表达,重组蛋白pNPC酶活为15.6U/L,最适反应温度为45℃,最适pH值为5.0,Mn2+对酶活力有明显的促进作用,SDS-PAGE表明重组蛋白分子量约为70kDa. |
| 关 键 词： | 康氏木霉 纤维二糖水解酶Ⅰ cbhI基因 克隆表达 |
Cloning of CBHI gene of Trichoderma koningii cellulase and expression in Escherichia coli |
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| HUANG Shihai,KANG Chao,HUANG Fei,CAO Xixiu,HE Xinping,WANG Sheng,WU Kongyang,CAO Pumei,LIANG Zhiqun,LI Xiangping. Cloning of CBHI gene of Trichoderma koningii cellulase and expression in Escherichia coli[J]. China Brewing, 2011, 0(2) | |
| Authors: | HUANG Shihai KANG Chao HUANG Fei CAO Xixiu HE Xinping WANG Sheng WU Kongyang CAO Pumei LIANG Zhiqun LI Xiangping |
| Affiliation: | HUANG Shihai1,KANG Chao1,HUANG Fei3,CAO Xixiu1,HE Xinping1,WANG Sheng1,WU Kongyang1,CAO Pumei 1,LIANG Zhiqun1,LI Xiangping2(1.College of Life Science and Technology,Guangxi University,Nanning 530005,China,2.Animal Reproduction Institute,3.Nanning New Technology Incubation Center,Nanning 530007,China) |
| Abstract: | Cellobiase cbhI gene was cloned with RT-PCR by using Trichoderma Koningi cDNA as template.The cbhI gene was inserted directly into the temperature-inducible prokaryotic expression vector pET-30a(+)and transformed into E.coli BL21(DE3) plysS strain after sequence.When induced with 4mmol/L IPTG for 4h,the BL21 strain produced a recombinant cellobiohydrolase.The results indicated that gene cbh I could expressed in strain BL21(DE3) plysS.Enzyme activity of pNPC was 15.6U/L under the conditions of temperature 45... |
| Keywords: | Trichoderma koningii cellobiohydrolase I gene cbhI cloning and expression |
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