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Nucleic Acids Research, 1984, Vol. 12, No. 4 2035-2046
© 1984

MOLECULAR BIOLOGY

Construction of a cDNA clone corresponding to mouse {alpha}1(IV) procollagen*

Cecilia L.S. dos Santos, Luisa Lina villa, Shigueko Sonohara+ and Ricardo Renzo Brentani

Ludwig Institute for Cancer Research Rua Prof. Antonio Prudente, 211-4°. andar (anexo), 01509, Säo Paulo +Laboratório de Oncologia Experimental, Faculdade de Medicina, Universidade de Säo Paulo Säo Paulo, Brazil

Received October 31, 1983. Revised February 2, 1984. Accepted February 2, 1984.

A new procedure for the synthesis of double stranded cDNA, based upon release of mRNA by "in vitro" translation, was used to clone type IV collagen. Collagen synthesizing polysomes selectively isolated from a mouse parietal yolk sac carcinoma (PYS-2) were used for translation in an heterologous cell-free system. Translation products were collagenase-sensitive and displayed an electrophoretic mobility correspondent to type IV collagen. Translation released mRNA was employed to construct a 100 base pairs long cDNA clone which hybridized to a 7,800 nucleotides long mRNA. Peptides synthesized by "in vitro" translation of hybrid selected mRNA displayed an electrophoretic mobility compatible with that of {alpha}1 (IV) collagen, were sensitive to collagenase and were immunoprecipitated by anti-type IV collagen antibody.


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