摘要
用Trizol试剂提取猪血白细胞总RNA,根据已发表的序列设计引物,采用RT-PCR方法扩增出猪β防御素-2(pBD-2)基因,并将该基因插入到真核表达载体pcDNA3.1A中,构建成猪β防御素-2基因的真核表达载体pcDNA3.1A/pBD-2,经磷酸钙介导转染HEK293T细胞进行表达。结果表明:本试验扩增的pBD-2基因与已发表的序列完全一致。表达产物经Western-blot检测,证明pBD-2能够在真核细胞HEK293T中表达。
Total RNA of porcine blood leucocytes was extracted by Trizol reagent. According to published eDNA sequence of porcine β- defensin - 2 （pBD - 2）, the eDNA encoding pBD - 2 was amplified by RT - PCR. Then the pBD - 2 gene was inserted into eukaryotic expression vector pcD- NA3.1A. The recombinant pcDNA3.1A- pBD - 2 plasmids were transfected into HEK293T cells mediated by calcium phosphate. Results showed that the sequence of pBD- 2 cDNA amplified was in consistence with that previously reported and pBD - 2 gene could be expressed in eukaryotie cells.
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2009年第1期78-81,共4页
Journal of Hebei Agricultural University
基金
2007年河北省科技支撑计划项目(07220401D)
河北农业大学校长基金项目
