CAJ | 학술논문

为建立延边黄牛白细胞介素2（IL-2）的基因克隆和序列分析的合理方法体系进行该研究．采集1月龄犊牛静脉血，用密度梯度离心法获取白细胞层，对白细胞进行总RNA的提取，并构建cDNA文库，根据Genbank牛IL-2基因序列（登录号：M12791．1）设计1对特异性引物，用反转录PCR（RT-PCR）技术扩增出目的基因片段，与克隆载体PMD18-T连接，并进行PCR和酶切鉴定，酶切结果为阳性的进行序列分析．结果表明：PCR扩增出大小为261bp的部分IL-2基因片断，与预期的片段大小一致；此序列与Genbank上牛IL-2从141～401bp片段的同源性为100％．
위건립연변황우백세포개소2（IL-2）적기인극륭화서렬분석적합리방법체계진행해연구．채집1월령독우정맥혈，용밀도제도리심법획취백세포층，대백세포진행총RNA적제취，병구건cDNA문고，근거Genbank우IL-2기인서렬（등록호：M12791．1）설계1대특이성인물，용반전록PCR（RT-PCR）기술확증출목적기인편단，여극륭재체PMD18-T련접，병진행PCR화매절감정，매절결과위양성적진행서렬분석．결과표명：PCR확증출대소위261bp적부분IL-2기인편단，여예기적편단대소일치；차서렬여Genbank상우IL-2종141～401bp편단적동원성위100％．
Studied the cloning and sequence analysis to build a reasonable method system of the gene. The test retrieval the total RNA from Yanbian calves WBC, reverse transcription to build cDNA library, according to IL-2 gene sequence enunciable in the Gen Bank to design a pair of specificity primer, use PCR technique to amplification purpose fragments, link cloning vectors PMD18-T, PCR and enzyme cuttings appraisement, sequence analysis of the masccline enzyme cuttings. The results showed that the size of PCR amplification IL-2 fragments is 261 bp,conformity for anticipant and coisogenic between cloning IL-2 gene sequences and Gen bank is 100%.