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Nucleic Acids Research, 1985, Vol. 13, No. 11 3917-3930
© 1985

Articles

Cloning of cDNA sequences for an Artemia salina hnRNP protein: evidence for conservation through evolution

Marilyn Cruz-Alvarez, Wlodzimierz Szer1 and Angel Pellicer

1Department of Biochemistry 550 First Avenue, New York University Medical Center, NY 10016, USA Department of Pathology and Kaplan Cancer Center 550 First Avenue, New York University Medical Center, NY 10016, USA

Received March 18, 1985. Revised May 14, 1985. Accepted May 14, 1985.

A cDNA clone was isolated for Artemia salina protein HD40, a component of heterogenous nuclear ribonucleoproteins. Enriched Artemia 15S poly(A)+ RNA was used as a template and double-stranded cDNA sequences were Inserted into the Pst I restriction endonuclease site of E. coli plasmid pBR322. Recombinant colonies were analyzed by positive hybrid selection of poly(A)+ RNA that directs the synthesis of protein HD40 In an 1n vitro assay. In vitro translation of the mRNA selected by recombinant clone 87HD yields a protein that is Immunopredpitated by anti-HD40 antibodies and that comigrates with authentic HD40 on gel electrophoresis. Partial proteolysis of protein HD40 and the in vitro translated product selected by clone 87HD produces the same peptide patterns. The size of the cloned insert 1s about 820 bp. The length of HD40 mRNA as determined by Northern blot analysis, Is about 1500 nucleotides. Southern blot analysis performed with DNA of different species (plant, avian, mammal) shows cross-hybridizing bands when probed with clone 87HD DNA suggesting that the HD40 gene 1s evolutionarily conserved


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