乙型肝炎与弓形虫复合DNA疫苗的构建与鉴定
Construction and identification of Complex DNA vaccine of hepatitis B and Toxoplasma gondii.
作者信息
Wei Qing-Kuan, Xiao Ting, Li Jin, Yin Kun, Jia Feng-Ju, Xu Chao, Zhao Gui-Hua, Cui Yong, Liu Gong-Zhen, Sun Hui, Jiang Hong-Tao, Yan Ge, Huang Bing-Cheng
机构信息
Shandong Academy of Medical Sciences, Shandong Institute of Parasitic Disease Jining 272033, China.
出版信息
Int J Clin Exp Med. 2015 Jun 15;8(6):9156-61. eCollection 2015.
OBJECTIVE
To construct and identify multi-gene recombinant expression vector pcDNA3-HBsAg-p30-ROP2.
METHOD
Primers were designed according to the gene sequences of restriction enzyme cutting site of recombinant pcDNA3-p30-ROP2 and hepatitis B surface antigen (HBsAg). The target fragment of HBsAg was amplified and cloned to expression vector pcDNA3-p30-ROP2 by restriction enzyme digestion and ligation. The recombinant expression vector pcDNA3-HBsAg-p30-ROP2 was identified by PCR detection, followed by enzyme restriction and sequencing.
RESULTS
The target fragment of HBsAg was successfully amplified, and the multi-gene eukaryotic expression vector pcDNA3-HBsAg-p30-ROP2 was established. PCR detection and restriction enzyme digestion showed that the length of the target fragment was consistent with the theoretical value. The recombinant expression vector contained the complete sequences of p30-ROP2 compound gene and HBsAg.
CONCLUSION
Multi-gene recombinant expression vector pcDNA3-HBsAg-p30-ROP2 was successfully established. The constructed expression vector could be used to develop multi-gene nucleic acid vaccines.
目的
构建并鉴定多基因重组表达载体pcDNA3-HBsAg-p30-ROP2。
方法
根据重组pcDNA3-p30-ROP2和乙型肝炎表面抗原（HBsAg）的酶切位点基因序列设计引物。通过酶切和连接将HBsAg的目标片段扩增并克隆至表达载体pcDNA3-p30-ROP2。通过PCR检测、酶切及测序对重组表达载体pcDNA3-HBsAg-p30-ROP2进行鉴定。
结果
成功扩增出HBsAg的目标片段，建立了多基因真核表达载体pcDNA3-HBsAg-p30-ROP2。PCR检测和酶切显示目标片段长度与理论值一致。重组表达载体包含p30-ROP2复合基因和HBsAg的完整序列。
结论
成功构建了多基因重组表达载体pcDNA3-HBsAg-p30-ROP2。构建的表达载体可用于开发多基因核酸疫苗。
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