变形链球菌葡聚糖结合蛋白C基因特异片段的分子克隆

The clonging and sequencing of a segment supposed to encode a glucan-binding domain of glucan-binding protein C of Streptococcus mutans

目的 :克隆变形链球菌葡聚糖结合蛋白C基因 (gbpC)编码序列的特异片段。方法 :根据文献报道的gbpC序列设计并合成一对寡核苷酸引物 ,PCR扩增位于gbpC编码序列 110 5～ 15 5 4bp间的一段特异片段 ,扩增产物经回收、酶切后 ,定向插入克隆载体puc18中 ,连接产物转化感受态大肠杆菌DH5α ,挑选阳性克隆 ,鉴定后进行序列测定。结果 :测序结果与Sato等报道的序列一致。结论 :成功克隆了gbpC基因片段 ,为进一步的研究 (探针标记、杂交及表达等 )

AIM: Cloning and sequencing of a segment supposed to encode a glucan-binding domain of glucan-binding protein C of Streptococcus mutans. METHODS: Design and synthesize a couple of oligodoxynucleotide primers in accordance with the sequence of gbpC of S.mutans 109cS reported by Sato et al , amplify a distinct segment of gbpC between 1105bp and 1554bp by polymerase chain reaction(PCR); after recovery and enzyme digestion, the PCR product was inserted into puc18 vector; the resulted plasmid was transformed into E.coli DH5α and the two-stranded DNA was sequenced after identification. RESULTS:The result of sequencing was consistent with that reported by Sato et al . CONCLUSION: The distinct segment of gbpC was cloned successfully and thus made a basis for further researches(probe marking, hybridization, expression et al ).