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| 白鹅催乳素基因的克隆及诱导表达条件的优化 | |
| 引用本文： | 郭丽,杨焕民,李鹏,康波.白鹅催乳素基因的克隆及诱导表达条件的优化[J].遗传,2008,30(11):1433-1438. |
| 作者姓名： | 郭丽 杨焕民 李鹏 康波 |
| 作者单位： | 1. 黑龙江八一农垦大学动物科技学院,大庆,163319 2. 吉林大学畜牧兽医学院,长春,130062 |
| 基金项目： | 黑龙江省科技厅科技攻关项目 |
| 摘 要： | 摘要: 运用RT-PCR方法, 从白鹅脑垂体总RNA中扩增得到了催乳素(Prolactin, PRL)基因编码区序列cDNA, 并将其克隆到pMD18-T载体上。DNA序列分析表明, PRL cDNA包括终止密码子在内的长度为690 bp，编码230个氨基酸残基的蛋白质, 与皖西白鹅的有所差异, 二者碱基同源性在99.57%, 氨基酸同源性达99.56%。将PRL基因编码区序列cDNA定向克隆到表达载体pET-32a (+)中, 构建表达质粒pET-32a(+)-PRL。该质粒的BL21 (DE3)转化菌在IPTG的诱导下可表达PRL基因融合蛋白, IPTG终浓度1 mmol/L, 37℃, 诱导4 h表达量最高, 表达量约占菌体总蛋白的28.96%。 |
| 关 键 词： | 白鹅 催乳素基因 克隆 表达 |
| 收稿时间： | 2008-01-20 |
| 修稿时间： | 2008-04-19 |
Cloning and optimizing prokaryotic induced expression conditions of prolactin in White Goose |
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| GUO Li,YANG Huan-Min,LI Peng,KANG Bo.Cloning and optimizing prokaryotic induced expression conditions of prolactin in White Goose[J].Hereditas,2008,30(11):1433-1438. | |
| Authors: | GUO Li YANG Huan-Min LI Peng KANG Bo |
| Affiliation: | 1. College of Animal Science and Veterinary Medicine, Heilongjiang August First Land Reclamation University, Daqing 163319, China; 2. College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China |
| Abstract: | Abstract: The mature segment gene of prolactin (PRL) in White Goose was amplified from pituitary by RT-PCR and then cloned into the pMD18-T vector. Sequencing analysis showed that the cDNA has a length of 690 bp including the termina-tion codon and encodes a protein composed of 230 amino acids, which differs from the published PRL cDNA sequence. There is a homology of 99.57% in base and 99.56% in amino acids with that of Wanxi White Goose, respectively. A pro-karyotic expression vector, pET-32a(+), was used to construct the recombinant plasmid pET-32a(+)-PRL to produce protein. Having been induced by IPTG，the host cell carrying the recombinant plasmid expressed the recombinant PRL. The opti-mal condition for expression is 1 mmol/L IPTG at 37℃. Based on this condition, the expression rose to the highest level by 4 h of induction, accounting for 28.96% of the total bacterial protein. |
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