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Partial characterization of guinea pig cholinephosphotransferase cDNA.

Abstract：

:An alternative molecular biology strategy is needed to characterize cholinephosphotransferase (CPT) gene because of the complexity of the problem associated with the solubilization of the membrane-bound enzyme without denaturation. We have synthesized five heterologous oligonucleotide probes based on the published yeast CPT gene sequence. Each probe (24 to 30 mers) was used as either forward or reverse flanking primers in combination with lambda gt11 primers to amplify a segment of DNA from a guinea pig liver 5'cDNA library by polymerase chain reaction (PCR). We detected several clones of varied size (0.1 kb to 2.2 kb) by subjecting the PCR products to 1.2% agarose gel electrophoresis. Southern blot of a 0.7 kb PCR product did hybridize with a 32P-labeled internal probe. Slot blot hybridization of guinea pig liver total RNA with the 32P-labeled 0.7 kb PCR product yielded positive transcripts with intensities proportional to the concentration of RNA. Furthermore, a 0.1 kb clone was sequenced and the observed sequence shared 96% homology with the yeast CPT gene sequence.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Das SK,Beg O,Mukherjee S

doi

10.1006/bbrc.1997.7862

subject

Has Abstract

pub_date

1997-12-18 00:00:00

pages

504-8

issue

2

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(97)97862-1

journal_volume

241

pub_type

杂志文章

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