title: Intron V, not intron I of human thrombopoietin, improves expression in the milk of transgenic mice regulated by goat beta-casein promoter.
abstract: Introns near 5' end of genes generally enhance gene expression because of an enhancer /a promoter within their sequence or as intron-mediated enhancement. Surprisingly, our previous experiments found that the vector containing the last intron (intron V) of human thromobopoietin (hTPO) expressed higher hTPO in cos-1 cell than the vector containing intron I regulated by cytomegalovirus promoter. Moreover, regulated by 1.0 kb rat whey acidic protein promoter, hTPO expression was higher in transgenic mice generated by intron V-TPOcDNA than in transgenic mice generated by TPOcDNA and TPOgDNA. However, it is unknown whether the enhancement of hTPO expression by intron I is decreased by uAUG7 at 5'-UTR of hTPO in vivo. Currently, we constructed vectors regulated by stronger 6.5 kb β-casein promoter, including pTPOGA (containing TPOcDNA), pTPOGB (containing TUR-TPOcDNA, TUR including exon1, intron I and non-coding exon2 of hTPO gene), pTPOGC (containing ΔTUR-TPOcDNA, nucleotides of TUR from uAUG7 to physiological AUG were deleted), pTPOGD (containing intron V-TPOcDNA) and pTPOGE (containing TPOgDNA), to evaluate the effect of intron I on hTPO expression and to further verify whether intron V enhances hTPO expression in the milk of transgenic mice. The results demonstrated that intron V, not intron I improved hTPO expression.
authors.affiliation: School of Biotechnology, Southern Medical University, North1838 Guangzhou Road, Guangzhou, 510515, PR. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Pediatrics, the Six Affiliated Hospital of Sun Yatsen University, 26 Yun Cun Erheng Road, Guangzhou, 510655, PR. China.
authors.forename: Hu
authors.initials: H
authors.lastname: Hao
authors.affiliation: School of Biotechnology, Southern Medical University, North1838 Guangzhou Road, Guangzhou, 510515, PR. China.
authors.forename: Mingqian
authors.initials: M
authors.lastname: Zhou
authors.affiliation: School of Public Health and Tropical Disease, Southern Medical University, North1838 Guangzhou Road, Guangzhou, 510515, PR. China.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Zhou
authors.affiliation: School of Biotechnology, Southern Medical University, North1838 Guangzhou Road, Guangzhou, 510515, PR. China.
authors.forename: Jianbin
authors.initials: J
authors.lastname: Ye
authors.affiliation: School of Biotechnology, Southern Medical University, North1838 Guangzhou Road, Guangzhou, 510515, PR. China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Ning
authors.affiliation: School of Biotechnology, Southern Medical University, North1838 Guangzhou Road, Guangzhou, 510515, PR. China.
authors.forename: Yunshan
authors.initials: Y
authors.lastname: Ning
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D019556:COS Cells; D002364:Caseins / Q000235:genetics* / Q000378:metabolism; D002522:Chlorocebus aethiops; D005260:Female; D006041:Goats; D006801:Humans; D007438:Introns; D008321:Mammary Glands, Animal / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008892:Milk / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D051381:Rats; D013926:Thrombopoietin / Q000235:genetics*
chemical_list: D020121:5' Untranslated Regions; D002364:Caseins; D013926:Thrombopoietin