CAJ | 학술논문

目的研究In-Fusion技术在HBV全进组克隆中的应用,并分析所获得的的序列信息.方法在浙江杭州地区收集23份慢性乙型肝炎患者血清,提取病毒核酸,分别设计PCR引物扩增病毒基因组和pMD18T载体,采用In-Fusion方法构建基因组测序质粒,并对序列进行分析.结果成功扩增出了23条HBV基因组序列,向NCBI提交的24株HBV全长序列,测序结果分析显示,B基因型11例,C基因型12例,没有B+C型,发现YMDD耐药株1例.结论 In-Fusion技术构建重组质粒是一种高效的连接方法,提高了连接率与成功率,可以大大简化目的片段与载体的连接,有效避免了在连接过程中常见的载体自连现象,探索了该技术在HBV序列研究中的初步应用.本研究丰富了我国的HBV全长基因组数据库,为HBV基因组特点、变异、药物、疫苗研究以及乙型肝炎的个体化治疗提供了参考依据.
목적 연구In-Fusion기술재HBV전진조극륭중적응용,병분석소획득적적서렬신식.방법 재절강항주지구수집23빈만성을형간염환자혈청,제취병독핵산,분별설계PCR인물확증병독기인조화pMD18T재체,채용In-Fusion방법구건기인조측서질립,병대서렬진행분석.결과 성공확증출료23조HBV기인조서렬,향NCBI제교적24주HBV전장서렬,측서결과분석현시,B기인형11례,C기인형12례,몰유B+C형,발현YMDD내약주1례.결론 In-Fusion기술구건중조질립시일충고효적련접방법,제고료련접솔여성공솔,가이대대간화목적편단여재체적련접,유효피면료재련접과정중상견적재체자련현상,탐색료해기술재HBV서렬연구중적초보응용.본연구봉부료아국적HBV전장기인조수거고,위HBV기인조특점、변이、약물、역묘연구이급을형간염적개체화치료제공료삼고의거.
Objective To investigate the use of In-Fusion technology in the sequencing of hepatitis B virus genome and analyzing the sequence of 23 strains of isolates.Methods Twenty-three strains of type B HBV genome were amplified by PCR from the serum collected in Zhejiang,China,and were inserted into the vector of pMD18T in the ways of In-Fusion technology,and the sequence of it was analyzed.Results The strains of the HBV isolates were types B and C,which were most common in China,and one isolate of YMDD mutation was found.Conclusion It was proven that the In-Fusion technology is an effective way to construct recombinant plasmid,avoiding self-connection of vector.The finding of this research provides a reference to the study of the genotype and evolution of hepatitis B virus in China.