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Nucleic Acids Research, 1983, Vol. 11, No. 17 6021-6039
© 1983

MOLECULAR BIOLOGY

Construction and characterization of type II collagen complementary deoxyribonucleic acid dones

Lewis N. Lukens, Anna Maria Frischauf*, Philip J. Pawlowski+,, George T. Brierley and Hans Lehrach*

Biology Department, Wesleyan University Middletown, CT 06457, USA *European Molecular Biology Laboratory Heidelberg, FRG

+Present address: Pfizer central Research, Groton, connecticut.

Received May 4, 1983. Accepted June 27, 1983.

The mRNA for type II collagen was purified from embryonic chick sternum or from purified sternal chondrocytes with guanidine thiocyanate as the extractant. Double-stranded cDNAs to procollagen mRNAs from sternum were synthesized and dC-tailed. After annealing with PatI-cleaved, dG-tailed pBR322, this DNA was used to transform Escherichia coli X1776. Transformed colonies were screened by colony hybridization to type I and II collagen cDNAs. Clones that preferentially hybridized to type II cDNA were characterized further. Four such cDNA clones, pCgII-2, 3, 10 and 12, with Inserts of 400, 320, 260 and 750 bp, have been identified as type II collagen cDNA clones by several criteria, including their preference for hybridizing with type II rather than type I collagen mRNAs in hybrid-selected translation experiments.


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