| 牛乳中乳酸乳球菌nisRK基因的克隆与序列分析 |
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| 引用本文： | 郝凤奇,王春凤,杨桂连,叶丽萍,杨中娜.牛乳中乳酸乳球菌nisRK基因的克隆与序列分析[J].吉林农业大学学报,2010,32(1):108-112. |
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| 作者姓名： | 郝凤奇 王春凤 杨桂连 叶丽萍 杨中娜 |
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| 作者单位： | 吉林农业大学动物科技学院,长春,130118 |
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| 基金项目： | 国家“863”计划项目(2007AA10Z322,2006AA10A205);;国家自然科学基金项目(30671573) |
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| 摘 要： | 以一株分离自牛乳中乳酸乳球菌染色体DNA为模板,采用PCR技术扩增出乳链菌肽生物合成的双组分调控元件(nisRK基因)。试剂盒回收纯化nisRK基因后将其克隆入以氨苄青霉素为选择压力的pGEM-T克隆载体中,再转化E.coliDH5α感受态细胞,筛选阳性克隆,提取重组体,进行酶切鉴定和PCR鉴定,并对nisRK基因进行序列测定,与已知序列进行同源性比较。结果表明成功地克隆出nisRK基因,全长为2 035 bp,与国外报道的nisRK基因同源性高达99.9%。
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| 关 键 词： | 乳酸乳球菌 nisRK基因 克隆 序列分析 |
| 收稿时间： | 2008-04-24 |
| 修稿时间： | 2008-07-01 |
Cloning and Sequence Analysis of nisRK Gene of Lactococcus lactis Isolated from Milk |
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| HAO Feng-qi,WANG Chun-feng,YANG Gui-lian,YE Li-ping,YANG Zhong-na.Cloning and Sequence Analysis of nisRK Gene of Lactococcus lactis Isolated from Milk[J].Journal of Jilin Agricultural University,2010,32(1):108-112. |
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| Authors: | HAO Feng-qi WANG Chun-feng YANG Gui-lian YE Li-ping YANG Zhong-na |
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| Institution: | College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118,China |
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| Abstract: | Using chromosome DNA of Lactococcus lactis isolated from milk as template, nisRK gene was amplified by PCR with Pfu DNA polymerase. After purification, the PCR product was cloned into pGEM-T vector containing ampicillin resistance gene to generate recombinant plasmid pGEM-T∶∶nisRK. Then pGEM-T∶∶nisRK was transformed into E.coli DH5α competent cells. Identification by restriction endonuclease digestion and PCR technique showed that the whole nisRK gene was cloned successfully. Sequence analysis indicated that nisRK gene, 2 035 bp, displayed 99.9% nucleotide homology with the published abroad. |
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| Keywords: | Lactococcus lactis nisRK gene cloning sequence analysis |
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