摘要
[目的]构建和表达人可溶性血管内皮生长因子受体2(VEGFR2)与白细胞介素12(IL-12)融合基因的真核表达质粒,并初步验证其抗肿瘤活性。[方法]根据Gen Bank公布的基因序列,设计并分别全基因合成全长人VEGFR2和IL-12基因,VEGFR2与IL-12通过Furin/2A(F2A)进行连接,融合基因克隆入真核表达载体p VAX1,构建p VAX1-VEGFR2-F2A-IL-12表达质粒。将p VAX1-VEGFR2-F2A-IL-12质粒瞬时转染293T细胞后,通过ELISA法测其真核表达能力。质粒注射B16荷瘤小鼠模型后,分析其抗肿瘤效果。[结果]质粒p VAX1-VEGFR2-F2A-IL-12构建成功;ELISA检测293T细胞转染上清液结果显示,VEGFR2和IL-12都能够在293T细胞中得到表达。质粒注射荷瘤小鼠模型后,显示出了较强的抑制B16肿瘤生长的能力,达到50%。[结论]p VAX1-VEGFR2-F2A-IL-12质粒成功构建和表达,该质粒具有较好的抑制B16肿瘤生长的能力,其抑制率达50%。
[ Objective ] To construct and express eukaryotic expression plasmid carrying the fusion gene of human VEGFR2 and IL - 12, and investigate its anti -tumor activity. [ Methods ] According to the gene sequence in the Genbank, design and synthe- size human VEGFR2 and IL- 12 gene. The VEGFR2 gene and IL- 12 gene were fusion by Furin/2A,and then the fusion gene was cloned into the eukaryotic expression vector pVAX1 to construct the expression plasmid pVAX1 - VEGFR2 - F2A - IL - 12. Then the recombinant expression plasmid was transferred into 293T cells. The expression of fusion gene was identified by ELISA. The anti -tumor effect was analyzed after the plasmid injected into the tumor bearing mice model. [ Results] The plas- mid pVAX1 - VEGFR2 - F2A - IL - 12 was successfully constructed. The expression of VEGFR2 gene and IL - 12 gene could be detected in the pVAX1 - VEGFR2 - F2A - IL - 12 transfected 293T ceils by ELISA. After injected into the tumor bearing mice model, the plasmid showed strong ability of inhibition tumor growth, up to 50%. [ Conclusion ] The eukaryotic expression plasmid pVAXI - VEGFR2 - F2A - IL - 12 is successfully constructed and expressed in 293T cells. The plasmid has the char-acteristics of inhibition of B16 tumor growth,and the inhibition ratio up to 50%.
出处
《生物技术》
CAS
CSCD
北大核心
2015年第4期311-314,390,共5页
Biotechnology
基金
黑龙江省中医管理局科研项目("二妙散对银屑病样小鼠的治疗作用及机制研究"
No.zhy12_w016)资助
