CAJ | 학술논문

为研究通过体外表达的方式获得具有生物学活性的禽流感 HA蛋白的方法，根据GenBank公布的禽流感H9亚型的基因序列设计引物，扩增出 H A基因全长片段，将扩增片段克隆到pFast-BacHTa载体上，构建重组杆粒rBacmid HA ，在脂质体介导下转染Sf9细胞制备重组杆状病毒，表达HA蛋白，并通过血凝抑制试验、Westernt blot、间接免疫荧光试验研究其生物学活性。结果显示，在Sf9细胞中成功表达了 HA蛋白，分子量约为65 kD ，且表达蛋白具有良好的生物学活性。
위연구통과체외표체적방식획득구유생물학활성적금류감 HA단백적방법，근거GenBank공포적금류감H9아형적기인서렬설계인물，확증출 H A기인전장편단，장확증편단극륭도pFast-BacHTa재체상，구건중조간립rBacmid HA ，재지질체개도하전염Sf9세포제비중조간상병독，표체HA단백，병통과혈응억제시험、Westernt blot、간접면역형광시험연구기생물학활성。결과현시，재Sf9세포중성공표체료 HA단백，분자량약위65 kD ，차표체단백구유량호적생물학활성。
This study intends to express HA protein of avian influenza in vitro ,and lay the foun-dation for the serological diagnosis method which distinguish avian influenza subtypes .The pri-mers were designed according to sequences of subtype H 9 avian influenza virus in GenBank ,am-plified fragment of HA gene ,and amplified fragments were cloned into pFastBacHTa vector .The recombinant plasmid was transformed into DH10 competent cells to produce recombinant bacmid rBacmid-HA ,recombinant baculovirus was obtained through the liposome mediated transfection of Sf9 cells ,and the target protein was expressed in the Sf9 cells ,the molecular weight of about 65 kD .By hemagglutination test ,Westernt-blot ,indirect immuno fluorescence test showed that the protein was successfully expressed ,and had a good biological activity .