CAJ | 학술논문

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.coli DH5α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%.
이일주분리자우유중유산유구균염색체DNA위모판,채용PCR기술확증출유련균태생물합성적쌍조분조공원건(nisRK기인).시제합회수순화nisRK기인후장기극륭입이안변청매소위선택압력적pGEM-T극륭재체중,재전화E.coli DH5α감수태세포,사선양성극륭,제취중조체,진행매절감정화PCR감정,병대nisRK기인진행서렬측정,여이지서렬진행동원성비교.결과표명성공지극륭출nisRK기인,전장위2 035 bp,여국외보도적nisRK기인동원성고체99.9%.
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 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.