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目的：克隆胃癌的相关基因。方法：提取人胃癌细胞总RNA，分离其mRN。以NatI／oligo（dT）12-18为引物合成双链。cDNA，除去小于400bp的cDNA片段后，连接EcoRⅠ人工接头，经NotⅠ酶酶切，插入定向克隆表达载体λExcellNotI／EcoRI／CIP，体外包装后转染大肠杆菌NM522，构建了人胃癌细胞MGC－803定向克隆表达。cDNA文库。结果：文库含1．2×106个重组子，重组率为96．7％。用PCR技术鉴定，文库中插入cDNA的平均大小为1kbc结论：该文库适于筛选各种丰度mRNA的cDNA克隆。
Objective: To clone genes related to gastric cancer. Methods: Total RNA of human gastric cancer cells was extracted and its mRN was isolated. Double strands were synthesized using Natl/oligo(dT)12-18 primers. After removing the cDNA fragment of less than 400 bp, it was ligated with EcoRI artificial linker, digested with NotI enzyme, inserted into directional cloning expression vector λExcellNotI/EcoRI/CIP, packaged in vitro and transfected into E. coli NM522 to construct human gastric cancer cell MGC-803. Clone expression. cDNA library. Results: The library contained 1.2 × 106 recombinants and the recombination rate was 96.7%. The average size of the inserted cDNA in the library was 1 kbc as determined by PCR. Conclusion: This library is suitable for screening cDNA clones of various abundance mRNAs.