[Molecular cloning and functional analysis of bovine mammary gland matrix attachment regions].
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 Primer5.0 and Vector7.0. 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 BE with Lipofectamine.Compared with control bovine ear fibroblast cells transfected with pEGFP-C1, the effect of cloned BMR was apparent in dispelling position effect and enhancing gene expression.
Yang
Rui-Feng
RF
Institute of Bio-Engineering, Northwest A & F University, Yang ling, Shaanxi 712100, China. ruifengy@163.com
Shu
Jian-Hong
JH
Li
Zhi-Yan
ZY
Liu
Jin-Long
JL
Zhang
Yong
Y
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000378:metabolism*; D045170:Matrix Attachment Regions / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D049452:Green Fluorescent Proteins