Cloning of bovine pyruvate carboxylase and 5' untranslated region variants

Animal Biotechnology
Cansu AgcaS S Donkin

Abstract

Bovine pyruvate carboxylase (PC; EC 6.4.1.1) cDNA was cloned by reverse transcription (RT) PCR. The coding region plus 3' untranslated region (UTR) of PC mRNA is 3926 bases and encodes 1178 amino acid PC precursor protein. A 5' rapid amplification of cDNA ends protocol was used to clone the 5' end of the mRNA. Six 5'UTR variants ranging from 68 to 363 bp were cloned. Bovine PC 5'UTR (bPC5') variants contain 68 (bPC5'A), 263 (bPC5'B), 363 (bPC5'C), 89 (bPC5'D), 275 (bPC5'E), and 178 bp (bPC5'F). All variants contain a common coding sequence. An RNase protection assay and RT-PCR analysis confirms the presence of the 5'UTR variants. The abundance of PC mRNA, determined by Northern blot analysis, indicates that PC is more abundant in gluconeogenic and lipogenic tissues where all PC variants are expressed compared with tissues that do not possess the full spectrum of PC transcripts. The data suggest that bPC5'A, bPC5'B, and bPC5'F are more abundant in bovine liver than the other variants.

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Citations

Dec 21, 2010·IUBMB Life·Jörg R AschenbachGregory B Penner
Apr 18, 2016·Journal of Dairy Science·C G WalkerJ R Roche
Mar 16, 2006·Journal of Dairy Science·J K DrackleyC K Reynolds
Jul 11, 2008·The Biochemical Journal·Sarawut JitrapakdeePaul V Attwood
May 2, 2021·Cancer & Metabolism·Violet A KieselDorothy Teegarden
Aug 5, 2008·Molecular Genetics and Metabolism·Dong WangDarryl C De Vivo

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Methods Mentioned

BETA
Biopsy
PCR
in vitro transcription
electrophoresis
Assay
X-ray

Software Mentioned

Align
Kodak 1D
BLAST
Kodak Digital Science
GeneStream
Translate
Expert Protein Analysis System Translate
Basic Local Alignment Search Tool ( BLAST )

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