【摘要】 ｍＲＮＡ差异显示技术可以显示ｍＲｗａ表达水平的差异。为了对用ｍｎｗＡ差异显示技术得到的差异条带进行研究，该文用改良碱裂解法抽提ｐＢＳ质粒，用ＥＣＯＲＶ将ｐＢＳ切成平头，利用ＴａｑＤＮａ聚合酶的非模板依赖性，在ｄｒｙＰ存在的条件下，用切成平头的ＰＢＳ质粒自制了Ｔ—Ａ克隆载体，对用ｍＲＮＡ差异显示方法得到的差异条带进行了重组，结果表明：用自制Ｔ一Ａ克隆载体对ＰＣＲ产物进行克隆的连接率较高，自制Ｔ一Ａ载体进行ＰＣＲ产物克隆是一简便可行的方法。更多还原

【Abstract】 The technique of mRNA differential display can show the changes of mRNA expression level. In order to clone theinteresting PAR bands produced by mRNA differential display, display Molecular cloning techinque was used in this protocal.pBS plasmid was extracted and restriction enzyme EcoR V was used to generate a blunt ends, and then T--A cloning vector was prepared in the presence of Taq DNA polymerase, dTTP and above pBS plasmid with a blunt ends. PCRproducts were cloned into the above ’T-A cloning vector, recombninant plasmid was analysed with EcoR and HindIII, a higher ligation efficency was generated. ’These results indicate that T-A cloning cloning technique producedpBSplasmid was single and this approach is suitable and reliable for PCR products cloning更多还原