NDLI: The last intron of the human thrombopoietin gene enhances expression in milk of transgenic mice

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The last intron of the human thrombopoietin gene enhances expression in milk of transgenic mice

Content Provider	Springer Nature Link
Author	Li, Yan Zhou, Mingqian Zhou, Hongwei Ning, Yunshan
Copyright Year	2013
Abstract	Introns can enhance gene expression levels. This effect is known as intron-mediated enhancement, which is different from that of enhancers or promoters. In our previous study, under the control of the cytomegalovirus or goat β-casein promoter, the vector containing intron V-TPOcDNA expressed the highest thrombopoietin (TPO) level, whereas the vector containing TPOgDNA expressed the lowest level. In order to verify whether intron V also improves TPO expression in the milk of transgenic mice, rat whey acidic protein promoter was used as regulatory element to construct mammary gland expression vectors including pTPOWA (containing TPOcDNA), pTPOWB (containing intron V-TPOcDNA), and pTPOWC (containing TPOgDNA). These vectors were transfected into HC-11 cells and the supernatants were analyzed at 48 h. The highest TPO level was found in pTPOWB (795 pg/mL) and the lowest level in pTPOWC (193 pg/mL). Then, corresponding vectors were microinjected into fertilized mice zygotes. Transgenic mice were identified by polymerase chain reaction and Southern blot. Enzyme-linked immunosorbent assay was performed to measure TPO levels in the milk of lactating transgenic mice. The highest and lowest TPO levels were found in transgenic mice carrying intron V-TPOcDNA (2,307 pg/mL) and in transgenic mice carrying TPOgDNA (242 pg/mL), respectively. Thus, intron V remarkably improved TPO expression in transgenic mice.
Starting Page	229
Ending Page	236
Page Count	8
File Format	PDF
ISSN	1438793X
Journal	Functional & Integrative Genomics
Volume Number	14
Issue Number	1
e-ISSN	14387948
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2013-11-28
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Thrombopoietin Intron V Whey acidic protein Transgenic mice Cell Biology Plant Genetics & Genomics Microbial Genetics and Genomics Biochemistry Bioinformatics Animal Genetics and Genomics
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine

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