CAJ | 학술논문

根据发表的猪圆环病毒Ⅱ型(PCV2)ORF2 基因序列,设计合成1对特异性引物,从分离到的PCV2吉林株(JL01)中扩增出PCV2 ORF2 579 bp的核苷酸片段,克隆到表达载体pET-32a中,经BamHⅠ和HindⅢ酶切及序列分析获得阳性重组表达质粒pET-32a-ORF2.将其转化到表达宿主菌BL21中,经IPTG诱导,成功表达了ORF2基因编码的部分结构蛋白,分子量为40 kD.通过SDS-PAGE 和Western检测表明,表达的重组蛋白能够被PCV2阳性血清所识别,具有良好的反应原性.
근거발표적저원배병독Ⅱ형(PCV2)ORF2 기인서렬,설계합성1대특이성인물,종분리도적PCV2길림주(JL01)중확증출PCV2 ORF2 579 bp적핵감산편단,극륭도표체재체pET-32a중,경BamHⅠ화HindⅢ매절급서렬분석획득양성중조표체질립pET-32a-ORF2.장기전화도표체숙주균BL21중,경IPTG유도,성공표체료ORF2기인편마적부분결구단백,분자량위40 kD.통과SDS-PAGE 화Western검측표명,표체적중조단백능구피PCV2양성혈청소식별,구유량호적반응원성.
Based on the published nucleotide sequence of ORF2 gene of porcine circovirus 2, a pair of primers were designed and synthesized. The 579 bp was amplified by polymerase chain reaction(PCR) from Jilin strain, and then the fragment was cloned into pET-32a vector, BamHI and Hind HI restriction endonuclease analysis and DNA sequencing were used to identify the recombinant plasmid of pET-32a-ORF2. A recombinant vector was transformed into E. Coli BL21 cells. Then about 40kD fusion protein was expressed in recombinant strain BL21 after being induced by IPTG.The expressed protein confirmed by SDS - PAGE and Western blotting analysis of reactionogenicity can react with the polyclonal antibody against PCV2.