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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 )奠定了基础
Objective: To clone a specific fragment of the gbpC coding sequence of Streptococcus mutans glucan binding protein C gene. Methods: A pair of oligonucleotide primers were designed and synthesized according to the reported gbpC sequences. A specific fragment located between 110 5 and 15 5 4bp of the coding sequence of gbpC was amplified by PCR. The amplified product was recovered, digested and inserted into Clone vector puc18, the ligation product transformed competent E. coli DH5α, positive clones were selected, identified and sequenced. Results: Sequencing results were consistent with those reported by Sato et al. Conclusion: The cloned gbpC gene fragment was successfully cloned, which laid the foundation for further research (probe labeling, hybridization and expression).