| 摘要: |
| ［摘要］ 目的 探讨通过体外扩增乙肝表面抗原（HBsAg）基因，构建真核表达载体pcDNA3-HBsAg-ROP2的可行性。方法 根据乙肝S基因序列及pcDNA3-p30-ROP2酶切位点等情况设计、合成引物，应用聚合酶链反应（PCR）技术，扩增HBsAg基因片段；应用回收、纯化、酶切、连接等技术，将HBsAg基因替换pcDNA3-p30-ROP2中的p30基因，并进行酶切、PCR扩增及测序鉴定。结果 PCR扩增出约0.7 kb的目的基因HBsAg，成功构建pcDNA3-HBsAg-ROP2重组载体。PCR、酶切结果与理论相符，重组体包含了HBsAg和ROP2基因的完整序列。结论 利用体外扩增乙肝表面抗原，可成功构建乙肝和弓形虫多功能重组载体pcDNA3-HBsAg-ROP2。 |
| 关键词: 乙肝表面抗原（HBsAg） 弓形虫 棒状体分泌抗原2（ROP2） 真核表达载体pcDNA3 基因重组 |
| DOI：10.3969/j.issn.1674-3806.2013.03.01 |
| 分类号:R 512.6+2 |
| 基金项目:山东省自然科学基金资助项目（编号：2009ZRC03083；2009ZRC03050）；山东省科技攻关项目（编号：2010GWZ20232）；济宁市科技发展计划项目（编号：济科2009-56-21） |
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| Construction and identification of multifunctional eukaryotic expression vector of HBsAg and ROP2 |
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WEI Qing-kuan, XIAO Ting，LI Jin,et al.
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Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease，Shandong Academy of Medical Science Key Laboratory of Molecular Immunology of Parasitic Disease，WHO Collaborating Centre for Lymphatic Filariasis and Taeniasis/Cysticercosis,Jining 272033，China
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| Abstract: |
| ［Abstract］ Objective To construct eukaryotic expression vector pcDNA3-HBsAg-ROP2 by amplifying HBsAg by PCR.Methods According to the specific sequence of HBsAg and restriction sites of pcDNA3-p30-ROP2, the primers were designed and synthesized. HBsAg was amplified by PCR, then purified HBsAg gene was inserted into vector instead of p30 by enzyme digestion and ligation, and the construct was identified by enzyme digestion, PCR and sequencing.Results 0.7 kb of HBsAg was aquired, and the pcDNA3-HBsAg-ROP2 eukaryotic expression recombinant plasmid was constructed.The results of PCR and digestion showed that HBsAg and ROP2 in the recombinant plasmid contained the complete genes sequences.Conclusion The recombinant plasmid pcDNA3-HBsAg-ROP2 was successfully constructed. |
| Key words: HBsAg Toxoplasma gondii Rhoptry protein 2(ROP2) pcDNA3 Gene recombinant |
