鸡IL-15基因的克隆及序列分析

Cloning and sequencing of the gene encoding chicken interleukin-15

目的利用基因工程的方法,将鸡白细胞介素15(ChIL-15)基因在原核表达系统中进行表达,为ChIL-15的应用提供科学依据。方法根据美国国家生物信息中心(NCBI)GenBank中已发表的ChIL-15基因,设计引物,用PCR方法扩增目的基因片段。结果成功从鸡的脾脏中扩增出含编码区的ChIL-15基因,ChIL-15基因为581bp,通过测序进行比对,表明ChIL-15基因克隆成功。提取的质粒经酶切鉴定、PCR鉴定、测序比对以及编码成氨基酸进行比对均与GenBank中登录号AF139097.1基本符合,核苷酸的同源性为99.82%。结论通过使用PCR方法成功扩增出来的ChIL-15基因片段,具有较高的同源性,并与已发表的基因序列相符。这为进一步研究ChIL-15基因的功能和应用提供了科学依据。

Objective To express the chicken interleukin-15(ChIL-15)gene in a prokaryotic expression system by means of genetic engineering to provide a scientific basis for the application of ChIL-15.Methods Primers were designed based on the CHER-15 gene in the GenBank of the National Center for Biological Information(NCBI),and PCR was utilized to amplify the target gene fragment.Result The ChIL-15 gene containing the coding region was successfully amplified from the spleen of chickens,with a length of 581 bp.Sequencing and comparison showed that the ChIL-15 gene was successfully cloned.The extracted plasmids were identified by enzyme digestion,PCR,sequencing,and amino acid coding and were consistent with the registration number AF139097.1 in GenBank.The nucleotide homology was 99.82%.Conclusion The ChIL-15 gene fragment successfully amplified by the PCR method has high homology and is consistent with the published gene sequence.This provides a scientific basis for further research on the function and application of the ChIL-15 gene.