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N-ras基因第12密码子、K-ras基因第12和13位密码子无已知的限制性内切酶的酶切位点,不能使用PCR-RFLP方法分析这些位点的突变。我们在PCR引物的3’端引入了一个误配的硷基使之正好成为某限制性内切酶的酶切位点,因而能使用PCR-RFLP方法分析上述位点的突变。本文使用PCR-RFLP方法分析了c-Ha-ras基因第12和61位、N-ras基因第12位、K-ras基因第12位和13位密码子的点突变。
The 12th codon of N-ras gene and the 12th and 13th codons of K-ras gene have no known restriction endonuclease sites and the mutations at these sites can not be analyzed by PCR-RFLP method. We introduced a mismatched base into the 3 ’end of the PCR primer to make it exactly the restriction site for a restriction enzyme, and therefore the PCR-RFLP method was used to analyze the above site mutations. In this paper, point mutations of 12th and 61th of c-Ha-ras gene, 12th of N-ras gene and 12th and 13th codons of K-ras gene were analyzed by PCR-RFLP method.