基因枪介导人精子顶体膜相关蛋白32基因疫苗在小鼠骨骼肌中的表达

Expression of gene vaccine of hSAMP32 in the mouse skeletal muscle medicated by gene gun

目的:观察pcDNA3.1(+)-人精子顶体膜相关蛋白32(hSAMP32)基因疫苗经基因枪介导肌肉接种后在小鼠体内的表达。方法:BALB/c雌性小鼠6只,实验组鼠(4只)经后腿股内侧肌以基因枪多点注射重组质粒pcDNA3.1(+)-hSAMP325μg,空载体对照组(1只)注射pcDNA3.1(+)载体5μg,空白对照组(1只)注射无菌生理盐水200μl。接种后第7d取小鼠股内侧肌注射部位肌肉,RT-PCR和原位杂交法检测目的基因mRNA的表达,以自制抗血清进行免疫组织化学染色检测目的基因蛋白的表达。取小鼠心、肝、脾、脑、肾、胚胎组织,PCR扩增,行基因疫苗的安全性检测。结果:RT-PCR和原位杂交检测结果显示实验组小鼠注射部位肌肉组织有目的基因及其蛋白的表达,对照组未见表达。实验组小鼠心、肝、脾、脑、肾、胚胎的基因组DNA均未扩增出目的基因条带,说明外源性基因没有整合到宿主染色体。结论:该基因疫苗可在小鼠骨骼肌内有效、安全地表达。

Aim： To observe the expression of gene vaccine of pcDNA3.1 （ ＋ ）-hSAMP32 in the mouse medicated by gene gun. Methods ： Six female BALB/C mice were used. 5 μg pcDNA3.1 （ ＋ ） -hSAMP32 was injected by gene gun into the hind leg vastus medialis of 4 mice（experimental group） , and 5 μg pcDNA3. 1 （ ＋ ） vector and 200 μl normal saline were injected into the other two mice respectively. The expressions of target gene mRNA and the target gene protein in the vastus medialis were detected by RT-PCR, in situ hybridization, and immunohistochemistry method at the 7th day after immunization. The tissue of heart,liver, spleen, kidney, brain and embry was obtained to evaluate the gene vaccine salty. Results： The expressions of target gene mRNA and the target gene protein in the vastus medialis of experimental group were positive. The target band was not found in the heart, liver, spleen, kidney, brain and embry tissue, indicating that hSAMP32 gene did not integrate into the host chromosome. Conclusion ： The gene vaccine of hSAMP32 can express in the mouse skeletal muscle safely and effectively.