【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%。更多还原

【Abstract】 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 lymphocytes 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.更多还原