摘要
目的 :探讨不同 β1,4半乳糖基转移酶 -V (β -1,4-GalTV)表达水平的生物学意义 ,构建正、反义 β -1,4-GalTV表达质粒。方法 :设计 β -1,4-GalTV全长引物 ;提取小鼠脑总RNA ,通过RT -PCR方法 ,得到全长 β -1,4-GalTV基因片段 ,将其克隆到 pGEM -T载体。再通过多克隆酶切位点 ,将 β -1,4-GalTV全长序列克隆到 pcD NA 3 .1表达载体中。设计反义引物 ,以pGEM -β -1,4-GalTV质粒为模板 ,将PCR产物克隆到 pcDNA 3 .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 ,得到了正、反义 β -1,4-GalTV表达质粒。 结论 :正、反义 β -1,4-GalTV表达质粒的构建 ,为进一步研究不同 β -1,4-GalTV表达水平的生物学意义奠定基础。
Objective:In order to study the biological effect of the expression level of β1,4-galactosyltransferase V (β-1,4-GalTV).Methods:The sense and anti-sense expression plasmids were constructed using molecular biological techniques. The whole gene sequence of β-1,4-GalTV 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-GalTV was cloned into the expression plasmid pcDNA3.1. With the anti-sense primers of β-1,4-GalTV, the anti-sense β-1,4-GalTV 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. Conclusions:The sense and anti-sense expression plasmids of β-1,4-GalTV were constructed in this experiment which will provide an approach to study the biological effect of different expression level of β-1,4-GalTV.
出处
《中国交通医学杂志》
2005年第1期5-7,共3页
Chinese Medical JOurnal of Communications
基金
江苏省高校高新技术产业发展项目(JH02-117)
江苏省高校自然科学研究项目(04KJB320114)
江苏省社会发展科技指导性计划项目(BS2004526)
