| 摘要: |
| 目的: 克隆人IL-12的cDNA。方法：利用RT-PCR从人NC 37淋巴母细胞中克隆出IL-12中两个亚基p35和p40的cDNA并进行序列测定。结果：从NC 37细胞中克隆的p35 cDNA序列(GenBank登录号为AF 101062)与先前登录在GenBank上的M65271 (NC 37细胞来源）和M 65291(RPMI 8866细胞来源,美国专利号：5648467）的p35 cDNA序列分别有2处和1处碱基不同：编码第44位氨基酸残基密码子的第3个碱基M 65291是C，而M 65271和本研究AF 101062的相应序列2是G；编码第244氨基酸残基密码子的第3个碱基，M 65271序列是C，而M 65291和AF 101062的相应序列是T；编码第247氨基酸残基密码子的第2个碱基M 65271序列是C，而M 65291和AF 101062的相应序列是T。此外，本研究克隆的p40 cDNA序列与登录在GenBank的p40 cDNA (M 38444)序列完全相同。结论：尽管本研究克隆的p35 cDNA序列与已知的两种p35 cDNA序列分别有2处和1处碱基不同，但其编码的氨基酸序列却与专利登记的p35 cDNA（M 65291）编码的氨基酸序列完全一致,表明克隆的IL-12基因所表达的蛋白质在理论上是符合目前药用标准的。 |
| 关键词: 白介素12 cDNA 聚合酶链式反应 |
| DOI： |
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| 基金项目: |
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| Cloning of human interleukin 12 cDNA |
| 郭亚军,卫立辛,王皓,卢洋,周倩,刘小萍,王华菁,刘彦君 |
| () |
| Abstract: |
| Objective:To clone human IL-12 cDNA. Methods: IL-12 cDNA (p35 and p40 cDNA) was cloned by RT-PCR from human lymphoblastoid B cell line NC 37, and their sequences were determined by automatic DNA sequencer. Results: There were 2 bases and one base different in p35 sequences of GenBank M 65271 and M 65291 (patent number: 5648467) respectively comparing to our p35 sequence (AF 101062). The third base of 44th code of M 65291 was C, but that of M 65271 or AF 101062 was G. The third base of the 244th code of M 65271 was C, but that of M 65291 or AF 101062 was T. The second base of 247th code was C, but that of M 65291 or AF 101062 was T. In addition, the sequence of our p40 cDNA was identical with that of GenBank (M 38444). Conclusion: Although there are 2 bases and one base different in p35 cDNA sequence of Genbank M 65271 and M 65291 respectively, comparing to AF 101062, the sequence of corresponding amino acids of AF 101062 is identical with that patented one. The recombinant IL-12 expressed by our p35 and p40 cDNA is acceptable in theory. |
| Key words: interleukin 12 cDNA RT-PCR |
