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Cloning and expression of fragment of the human ALEX2 cDNA

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Author:
No author available
Journal Title:
JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
Issue:
3
DOI:
10.3321/j.issn:1000-2790.2004.03.002
Key Word:
ALEX2;克隆;原核表达

Abstract: 目的: 克隆人ALEX2基因部分片段的cDNA,构建重组表达载体并诱导其表达. 方法: 通过RT-PCR技术从人胚胎脑组织中特异扩增出ALEX2基因部分编码序列片断(630~1058位核苷酸),克隆入pGEM-T easy载体中,测序后亚克隆于表达载体pGEX-4T-3中,酶切鉴定正确,转化大肠杆菌DH5α,经IPTG诱导4 h后,可表达Mr417 000的目的蛋白. 结果: 特异扩增出大小约440 bp的片断,经测序与已知序列完全一致,构建了融合蛋白的重组表达质粒PGEX-ALEX2,表达的融合蛋白产物经凝胶薄层扫描检测,占菌体总蛋白量的15.5%. 结论: 获得了人ALEX2部分基因编码片断及其原核表达产物,为下一步ALEX2 mAb的制备和研究其生物学功能奠定了基础.

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