原生动物Δ4-脂肪酸去饱和酶基因在卵巢细胞中的表达  

Protozoan Δ4-fatty acid desaturase gene expression in CHO cells

作　　者：汪坤福[1] 朱贵明[1] 张涛[1] 孙洁[1] 江旭东[1] 王明富[1] 

机构地区：[1]佳木斯大学,黑龙江佳木斯154007

出　　处：《中国老年学杂志》2013年第13期3091-3092,共2页Chinese Journal of Gerontology

基　　金：国家自然科学基金(No.310000984);黑龙江省教育厅项目(No.12511563);黑龙江省研究生创新基金(No.YJSCX2012-367HLJ)

摘　　要：目的探讨优化、合成原生动物Δ4-脂肪酸去饱和酶基因,并构建其哺乳动物表达载体,将基因导入卵巢细胞(CHO)中进行初步表达。方法目的基因经优化后人工合成全长编码区,利用双酶切连接法将其连入哺乳动物表达载体pcDNA3.1(-)。采用脂质体法稳定转染CHO细胞后应用PCR和RT-PCR法检测Δ4-脂肪酸去饱和酶基因的整合和表达情况。结果成功构建了Δ4-脂肪酸去饱和酶基因的表达载体pcDNA3.1-D4,并整合进CHO细胞基因组,实现了该基因在CHO中的表达。结论原生动物Δ4-脂肪酸去饱和酶基因哺乳动物表达载体的成功构建以及Δ4-脂肪酸去饱和酶基因表达后的分析为下一步深入研究Δ4-脂肪酸去饱和酶基因的功能和转基因动物的制作打下了基础。Objective To combine and optimize the gene of protozoa Δ4-fatty acid desaturase and construct its expression vector in mammalian,then introduce into CHO cells for preliminary expression analysis.Methods The optimized full-length coding region of this target gene was synthesised,be connected to the mammalian expression vector pcDNA3.1（-） by double digestion connection.After using the liposome method to stably transfect CHO cells,Δ4-fatty acid desaturase gene integration was detected.Results The Δ4-fatty acid desaturase enzyme gene expression vector pcDNA3.1-D4 was successfully constructed,integrated it into the CHO cell genome,and expressed the gene in CHO.Conclusions Protozoa Δ4-fatty acid desaturase gene mammalian expression vector is successfully constructed and laid a foundation on the aspect of further in-depth study for Δ4-fatty acid desaturase gene function and the production of transgenic animals through the Δ4-fatty acid desaturase gene expression analysis.

关 键 词：D4基因 表达载体 二十二碳六烯酸(DHA) 中国仓鼠卵巢细胞 

分 类 号：R730.59[医药卫生—肿瘤]