Synthesis, molecular cloning, and restriction analysis of DNA complementary to vitamin D-dependent calcium-binding protein mRNA from rat duodenum

C. Desplan, M. Thomasset, M. Moukhtar

Research output: Contribution to journalArticle

Abstract

The mRNA coding for rat intestinal calcium-binding protein, a vitamin D3-induced protein (M(r) 7500), has been partially purified from growing rat duodenum. Double-stranded DNA synthesized from the purified mRNA preparation was inserted into the PstI site of pBR322, using the oligo(dG-dC) tailing procedure. Clones containing DNA complementary to vitamin D-dependent calcium-binding protein mRNA were selected by differential colony hybridization with [32P] cDNA synthesized from enriched or low vitamin D-dependent calcium-binding protein mRNA preparations. Plasmid DNAs from the selected clones were each verified by both a solution hybrid-arrest assay and a filter hybrid-selection assay. Four recombinant clones showed identical endonuclease restriction maps and contained inserts ranging from 250 to 380 base pairs.

Original languageEnglish (US)
Pages (from-to)2762-2765
Number of pages4
JournalJournal of Biological Chemistry
Volume258
Issue number5
StatePublished - 1983

Fingerprint

S100 Calcium Binding Protein G
Cloning
Molecular Cloning
Duodenum
Rats
Complementary DNA
Messenger RNA
Clone Cells
Assays
Calcium-Binding Proteins
Cholecalciferol
DNA
DNA Restriction Enzymes
Tailings
Base Pairing
Plasmids
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Synthesis, molecular cloning, and restriction analysis of DNA complementary to vitamin D-dependent calcium-binding protein mRNA from rat duodenum. / Desplan, C.; Thomasset, M.; Moukhtar, M.

In: Journal of Biological Chemistry, Vol. 258, No. 5, 1983, p. 2762-2765.

Research output: Contribution to journalArticle

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