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Construction and identification of colorectal carcinoma cDNA phage expression library
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 18
- DOI:
- 10.3321/j.issn:1000-2790.2003.18.002
- Key Word:
- 结肠直肠肿瘤;噬菌体;cDNA;文库
Abstract： 目的: 构建人源性大肠癌抗原cDNA噬菌体表达文库. 方法: 从人大肠癌组织中提取总RNA,纯化mRNA,用RT-PCR反转录合成cDNA第一链,用LD-PCR合成cDNA第二链,除去小于500 bp的小片段,与去磷酸化的λTriplEx2噬菌体连接,体外包装. 转染E.coli XL1-Blue大肠杆菌,滴定文库的滴度,确定文库容量大小,用ITPG和X-gal测定重组率. 用SfiⅠ酶切鉴定插入的cDNA片段大小. 结果: 构建成含2.39×106重组子的人大肠癌抗原cDNA噬菌体表达文库,重组率为97.5%,插入外源cDNA片段大小范围从1～4 kb,平均长约2.4 kb. 结论: 成功构建人大肠癌抗原cDNA噬菌体表达文库,适合于大批量筛选cDNA克隆的大肠癌相关抗原基因.
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