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%。
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 en- donuclease 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.
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2010年第1期108-112,共5页
Journal of Jilin Agricultural University
基金
国家"863"计划项目(2007AA10Z322
2006AA10A205)
国家自然科学基金项目(30671573)
