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RT-PCR克隆人IL-6、IL-10、IL-13和SCF等四种细胞因子膜外区cDNA 被引量:1

Cloning of cDNAs Encoding Extra-cellular Domain of Human IL-6, IL-10, IL-13 and SCF
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摘要 应用RT—PCR方法,体外扩增获得人IL—6、IL—10、IL—13和SCF膜外区蛋白质编码序列.又运用基因重组手段,将这些基因序列分别插入到表达性载体PBV220中,转化大肠肝菌DH5α,经过筛选和鉴定,获得了上述细胞因子膜外区cDNA克隆.SDS—PAGE和生物学活性测定证实:IL—6、IL—10和IL—13已在大肠肝菌中得到表达,尤其是IL—6获得了高效表达,其表达量约占菌体总蛋白质含量的30%左右. Human IL-6,IL-10,IL-13 and SCF cDNAs encoding extra - cellular domain were isolated by reverse transcription polymerase reaction (RT-PCR). These cDNA fragments were inserted into expression vector PBV220 which contains PRP_(L), promoters and SD sequence. Then the E. coli DH5α were tronsformed with the recombinant plasmids. After screening, the clones carrying those recombinant plasmids were identified. SDS-PAEG and biological assay indicated that human IL-6, IL-10 and IL-13 were expressed in E.coli. The expression level of rhIL-6 was as high as 30% of the total bacterial protein.
出处 《中国肿瘤生物治疗杂志》 CAS CSCD 1995年第1期73-75,共3页 Chinese Journal of Cancer Biotherapy
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