NDLI: Molecular analysis of the first intron in the bovine myostatin gene

Content Provider	Springer Nature Link
Author	He, Yulong Wu, Yuehong Lan, Zhigang Liu, Yonggang Zhang, Yong
Copyright Year	2010
Abstract	To study the mechanism of transcription and expression of the myostatin gene, we cloned and analyzed the sequence of the bovine myostatin gene promoter and first intron from Qinchuan and Red Angus cattle, then constructed eukaryotic expression vectors encoding the GFP vector by replacing the CMV promoter with the bovine myostatin promoter using PCR method, thereby obtaining an expression vector coding GFP report gene with first intron (identified as pEGFP-MSTNPro-intron1). By transfecting C2C12 cells with the vectors, we then compared the effect on GFP gene expression of the promoter and normal first intron of Qinchuan and Red Angus cattle with that from the promoter and a Qinchuan allele with a 16 base pair insertion. After 48 h incubation, fluorescent indices (FIs), which indicate the expression rate and intensity of gene GFP expression, were analyzed by flow cytometry (FCM). Results showed that Qinchuan sequence homology of promoter was 99% with Red Angus, that Qinchuan first intron sequence homology was 99.51% with Red Angus and that first intron homologies of Qinchuan and Red Angus were 99.08 and 99.02%, respectively, with Accession No.AF320998 in GenBank. Expression of the GFP gene did not differ significantly between preparations using the Qinchuan versus Red Angus promoter. Preparations with a construct that included the first intron had higher GFP gene expression in C2C12 cells than those whose construct lacked the first intron (P < 0.05 or P < 0.01). However, there was no significant difference (P > 0.05) in gene expression between normal first intron and 16 bp insertion first intron (+16 bp) preparations.
Starting Page	4643
Ending Page	4649
Page Count	7
File Format	PDF
ISSN	03014851
Journal	Molecular Biology Reports
Volume Number	38
Issue Number	7
e-ISSN	15734978
Language	English
Publisher	Springer Netherlands
Publisher Date	2010-12-02
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Myostatin Promoter First intron C2C12 cell line Bovine Animal Anatomy / Morphology / Histology Animal Biochemistry
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine Molecular Biology

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