Title

牛乳腺核基质附着区的克隆及其功能研究

Translated Titles

Molecular Cloning and Functional Analysis of Bovine Mammary Gland Matrix Attachment Regions

Authors

杨瑞锋(Rui-Feng Yang);舒建洪(Jian-Hong Shu);李志艳(Zhi-Yan Li);刘金龙(Jin-Long Liu);张涌(Yong Zhang)

Key Words

核基质附着区 ; 克隆 ; 细胞表达 ; matrix attachment regions ; clone ; cell expression

PublicationName

遺傳

Volume or Term/Year and Month of Publication

28卷1期(2006 / 01 / 01)

Page #

31 - 35

Content Language

簡體中文

Chinese Abstract

从奶牛的全血中提取基因组DNA,参照GenBank已有序列,通过Primer5.0和Vector 7.0辅助设计一对引物,克隆了牛乳腺核基质附着区(BMRs)。利用生物学软件对其进行初步分析后,TA克隆于pMD 18-T vector上。上下游引物5'端分别引入Kpn2 I和Xho I酶切位点,将所得的BMRs克隆到pEGFP-C1的报告基因下游,构建成表达载体pBE,脂质休法转染牛耳成纤维细胞。以转染pEGFP-C1的牛耳成纤维细胞作为对照,发现所克隆的BMRs功能明显,在消除位置效应、增强报告基因表达方面具有一定的作用。

English Abstract

The bovine genomic DNA was extracted from bovine blood, then bovine mammary gland matrix attachment region (BMARs) was cloned using a pair of primers, which were designed based on the related sequences in GenBank through bio-software Primer 5.0 and Vector 7.0. Upon preliminary analysis with bio-software, BMARs was TA cloned into pMD 18-T vector. By means of adding Kpn2 I and Xho I to 5' upstream of sensitive and antisensitive primers respectively, expressing vector BE was constructed after BMAR was cloned into the downstream of the reporter gene in pEGFP-C1. Bovine ear fibroblast cells were transfected by expressing vector pBE with Lipofectamine. Compared with control bovine ear fibroblast cells transfected with pEGFP-8C1, the effect of cloned BMR was apparent in dispelling position effect and enhancing gene expression.

Topic Category 醫藥衛生 > 醫藥總論
醫藥衛生 > 基礎醫學
生物農學 > 生物科學