title: [Cloning and functional verification of U6 and 7SK promoter of small RNA from Bama mini-pig in Guangxi].
abstract: To investigate the functions of U6 and 7SK of Bama mini-pig and produce Bama mini-pig with silenced GGTA1 gene, the siRNA promoters U6 and 7SK were cloned, ligated into pMD18-shEGFP, and co-transfected with PEGFP- N1 into PK-15 kidney cells of pigs to be used in RNAi experiments. The functions of the two promoters in pig cells were verified using pMD18-hU6-shEGFP as the positive control, pMD18-shEGFP vector without promoter as the negative control, PEGFP-N1 as the first blank control, ddH2O in replacement of the plasmid as the second blank control. The results showed that the lengths of U6 and 7SK in Bama mini-pig were 553 bp and 437 bp, respectively. Vectors pMD18-pU6- shEGFP and pMD18-p7SK-shEGFP were constructed and transfected into PK-15 cells from pigs. Promoters pU6 and p7SK proved to express high levels of siRNA activity and can be used in the experiment of silencing α-1,3galactosyltransferase gene.
authors.affiliation: College of Animal Science and Technology, Guangxi University, Nanning 530005, China. 410537678@qq.com
authors.forename: Shi-Jin
authors.initials: SJ
authors.lastname: Chen
authors.forename: Jing
authors.initials: J
authors.lastname: Fan
authors.forename: Qin-Yang
authors.initials: QY
authors.lastname: Jiang
authors.forename: Gan-Qiu
authors.initials: GQ
authors.lastname: Lan
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Guo
authors.forename: Ya-Fen
authors.initials: YF
authors.lastname: Guo
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000502:physiology*; D012320:RNA Polymerase III / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D012320:RNA Polymerase III