中国人白细胞介素-12(hIL-12)cDNA克隆及序列分析  

Cloning and Sequence Analysis of Chinese Interleukin-12(hIL-12) cDNA

作　　者：韩红兵[1,2] 徐曙光[1,2] 陆泉枝[1,2] 连正兴[1,2] 田亮[1,2] 潘求真[1,2] 李宁[3,2] 

机构地区：[1]中国农业大学动物科技学院 [2]中国农业大学农业部畜禽遗传育种重点实验室,北京100094 [3]中国农业大学农业生物技术国家重点实验室

出　　处：《农业生物技术学报》2005年第4期456-460,共5页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.30170675)资助。

摘　　要：根据已发表的IL-12两个亚基P35cDNA序列(NM-000882)与P40cDNA序列(NM-002187)设计2对引物。以LPS刺激的人脐血淋巴细胞为材料,采用RT-PCR技术从总RNA扩增hIL-12基因,包括完整的前体蛋白编码序列。所得到的序列与GenBank中发表的一致,但与CLMF和NKSF序列有所差异。P35同CLMF相比,244aa密码子GAC(Asp)→GAT(Asp)、247aa密码子ACG(Thr)→ATG(Met);与NKSF比较,44aa密码子GTC(Val)→GTG(Val)。P40同NKSF比较239aa密码子AAC(Asn)→AAG(Lys),291aa密码子ACC(Thr)→ACG(Thr);P40同CLMF相比较,氨基酸与核苷酸序列完全一致。通过与不同物种氨基酸及核苷酸序列比较分析,P40亚基与其它动物的同源性80%以上,P35亚基与其它动物同源性在70%。Based on the published human L-12 （hIL-12） cDNA sequences in GenBank, two sets of primers were designed and synthesized. The total RNA was extracted from LPS-stimulated lymphoeytes of cord blood of a Chinese newborn, cDNAs of P40 （1100 bp） and P35 （870 bp） subunits of hIL-12 were amplified by RT-PCR and cloned into pMD18- T vector respectively. Comparison of the acquired sequence with submitted sequences in the GenBank by the BLAST showed that the sequences were identical to that published in GenBank. But difference does exist when compared to sequences of CLMF and NKSF. The P35 sequence is different from that of CLMF, in which 244 amino acid cordon changes from GAC（Asp） to GAT（Asp）, 247 amino acid cordon changes from ACG（Thr） to ATG（Met）, comparing with NKSF, 44 amino acid cordon changes from GTC（Val） to GTG（Val）. Amino acid and nucleotide sequences of the P40 subunit are identical with that of CLMF. However there were several differences between P40 and NKSF： 239 amino acid cordon changes from AAC（Asn） to AAG（Lys）, 291 amino acid cordon changes from ACC （Thr） to ACG（Thr）. Sequence analysis showed that the cloned sequence of p40 subunit had a similarity over 80% and the P35 subunit over 70% in amino acid and nucleotide sequences compared with other animals.

关 键 词：IL-12基因 cDNA克隆 序列分析 人白细胞介素 RT-PCR技术 CDNA序列 GENBANK 核苷酸序列 中国 IL-12 

分 类 号：Q78[生物学—分子生物学] Q26