摘要
目的 :构建 β- 1,4 -半乳糖基转移酶 - V(β- 1,4 - Gal TV)正、反义表达质粒 ,并将其转染到人脑星形胶质细胞瘤细胞 SHG- 4 4中 ,为探讨神经胶质瘤细胞表达β- 1,4 - Gal TV的生物学意义提供研究基础。方法 :构建 β- 1,4 - Gal TV表达质粒 ,并将其转染到人脑星形胶质细胞瘤细胞 SHG- 4 4中 ;用 Western印迹和 Northern印迹鉴定转染情况。结果 :通过酶切鉴定和测序分析 ,实验成功构建 β- 1,4 - Gal TV表达质粒。将构建的表达质粒转染到胶质细胞瘤细胞 SHG- 4 4中 ,经鉴定表明 ,转染的正义表达质粒能增加胶质细胞瘤细胞 SHG- 4 4中的 β- 1,4 -Gal TV表达量 ,而转染反义质粒则能降低细胞中 β- 1,4 - Gal TV表达量。结论 :正、反义 β- 1,4 - Gal TV表达质粒在人脑星形胶质细胞瘤细胞 SHG- 4 4中的稳定转染和表达 ,对分析胶质瘤细胞表达β- 1,4 - Gal TV的意义如对肿瘤细胞的增殖。
Objective:In order to study the biological effects of β-1,4-galactosyltransferase V (β-1,4-GalTV) expressed in astrocytoma,the sense and antisense expression plasmids of β-1,4-GalTV were constructed and transfected into SHG-44 cells, one kind of astrocytoma cell lines in this study.Methods: The whole gene sequence of β-1,4-GalTV was cloned into the pGEM-T vector and subcloned into the expression plasmid pcDNA3.1.In order to construct the antisense plasmid of β-1,4-GalTV, the fragment from the start code ATG of β-1,4-GalTV was generated by PCR by using the antisense primers.The plasmid containing sense or antisense β-1,4-GalTV was verified by EcoR I / Xho I digestion and DNA sequencing.The constructed expression plasmids were transfected into SHG-44 cells and certificated by Western Blot and Northern Blot analysis.Results:It was found that the sense and antisense expression plasmids of β-1,4-GalTV were constructed and transfected into the SHG-44 cells successfully.Conclusion:The astrocytoma cells transfected with the expression plasmids of β-1,4-GalTV will help to study the effects of β-1,4-GalTV on proliferation, adhesion and migration of astrocytoma cells.
出处
《南通大学学报（医学版）》
2005年第1期7-9,12,共4页
Journal of Nantong University(Medical sciences)
基金
江苏省自然科学资金资助项目 ( BK2 0 0 3 0 3 5 )
江苏省高校自然科学研究项目 ( 0 3 KJB180 10 9)
江苏省社会发展科技指导性计划项目 ( BS2 0 0 45 2 6)
