基因群体遗传多态性检测方法的比较研究

Comparative Study on Detecting Methods for the Genetic Polymorphism of Gene in Population

[目的]寻找基因的群体遗传多态性检测的理想方法,为基因突变的群体检测提供方法指导。[方法]以104头中国荷斯坦牛群为研究对象,扩增了牛TLR4基因243 bp特异性片段,分别采用SSCP和创造酶切位点的RFLP(CRS-RFLP)方法检测了扩增片段的第27 bp处C→T的突变,并对其方法进行了比较分析。[结果]结果表明,在104头个体的扩增片段中,SSCP未能检测出多态性,而CRS-RFLP方法检测出了多态性。[结论]在不能直接采用RFLP方法的情况下,与SSCP相比,创造酶切位点RFLP(CRS-RFLP)方法为群体多态性检测的理想选择。

[ Objective] This experiment was conducted to search the optimal methods for detecting gene poiymorphism in populations and to provide guidance for population detection of gene mutation. [ Method] 243 hp fragments of TLR4 gene for 104 Chinese Holstein were amplified by polymerase chain reaction, in which it had a mutation in the 27th base. The mutation was detected using Single-Strand ConformationPoly- morphism （SSCP） created restriction site combined restriction fragment poiymorphism（RCS-RFLP） respectively. The advantages and disadvantages of the two methods were analyzed. [ Result] The results showed that the mutation in this study could be detected easily and precisely by CRS-RFLP, but it could not be detected by SSCP in any condition. [ Conclusion] Compared with SSCP, CRS-RFLP was the Optimal method for gene polymorphism in populations.