摘要
目的；通过构建人扁桃体淋巴细胞ｃＤＮＡ质粒重组体，为深入研究淋巴细胞的生物学功能奠定实验基础。方法：采用ＤＮＡ重组技术，以人新鲜扁桃体淋巴细胞为实验材料从中提取ｍＲＮＡ，以ｍＲＮＡ为模板合成ｃＤＮＡ，然后与质粒ｐＳＰＯＲＴＩ进行定向连接并转入到Ｅ．ｃｏｌｉＪＭ１０９中进行了分子克隆。结果：阳性克隆玷９０％，阴性克隆占１０％，转化率为１．３５×１０＾４克隆／微升连接体系，插入片段长度为０．５－２．
Objective: To lay a foundation for the test of biochemistry mechanism of the lymph9cytes, we constructed of plasmid recombinant of the human tomsil lymphocytes cDNA. Method: DNA recombinant technique was used. The lymphocytes of the human fresh tonsils were used as the materials for the test and mRNA was purified from the materials, then mRNA being the template, cDNA was synthesized. The fragments of cDNA were inserted into the plasmid pSPORT I in only one orientation to form recombinant DNA molecules, which were then introduced to E. coli JM 109. Result:90 percent were positive clones and 10 percent were negative therefore, the transformation efficiency of each microliter ligation reactions was 1.35 × 104 clones. The length of fragments inserted was from 0. 5 to 2.3Kb, average value 1.3Kb. Conclusion: Plasmid recombinant of the human tonsil lymphocytes cDNA has been successfully established.
关键词
扁桃体
淋巴细胞
重组DNA
质粒
分子克隆
human tonsil lymphocytes
recombinant DNA
Plasmid
molecules clone
transformation
