【摘要】 目的 :为了探讨不同 β1,4半乳糖基转移酶 - I(β1,4 - galactosyltransferase I,β- 1,4 - Gal T- 1)表达水平的生物学意义。方法 :通过分子生物学手段 ,构建正、反义β- 1,4 - Gal T- I表达质粒。:设计β- 1,4 - Gal T- I全长引物 ;提取小鼠脑总 RNA,通过 RT- PCR方法 ,得到全长 β- 1,4 - Gal T- I基因片段 ,将其克隆到 p GEM- T载体。再通过多克隆酶切位点 ,将β- 1,4 - Gal T- I全长序列克隆到 pc DNA3.1表达载体中。设计反义引物 ,以 p GEM- β- 1,4 - Gal T- I质粒为模板 ,将 PCR产物克隆到 pc DNA3.1表达载体中。通过酶切和测序鉴定构建的质粒。结果 :通过酶切和测序证实 ,实验得到了正、反义β- 1,4 - Gal T- I表达质粒。结论 :正、反义β- 1,4 - Gal T- I表达质粒的构建 ,为进一步研究不同 β- 1,4 - Gal T- I表达水平的生物学意义奠定基础更多还原

【Abstract】 objective:In order to study the biological effect of the expression level of β1,4-galactosyltransferase I(β-1,4-GalT-I).Method:The sense and anti-sense expression plasmids were constructed using molecular biological techniques. The whole gene sequence of β-1,4-GalT-I was obtained by RT-PCR from total RNA of mouse brains and cloned into the pGEM-T vector. Utilizing the MCS(multi-clonal site),the whole gene sequence of β-1,4-GalT-I was cloned into the expression plasmid pcDNA3.1. With the anti-sense primers of β-1,4-GalT-I, the antisense β-1,4-GalT-I was obtained and cloned into the expression plasmid pcDNA3.1 by PCR. Results:Certificated by restriction enzymes digestion and DNA sequencing, the sense and anti-sense expression plasmids were constructed successfully. Conclusion:The sense and anti-sense expression plasmids of β-1,4-GalT-I were constructed in this experiment which will provide an approach to study the biological effect of different expression level of β-1,4-GalT-I.更多还原