Isolation of full-length putative rat lysophospholipase cDNA using improved methods for mRNA isolation and cDNA cloning

Biochemistry. 1987 Mar 24;26(6):1617-25. doi: 10.1021/bi00380a020.

Abstract

We have cloned a full-length putative rat pancreatic lysophospholipase cDNA by an improved mRNA isolation method and cDNA cloning strategy. These new methods allow the construction of a cDNA library from the adult rat pancreas in which the majority of recombinant clones contained complete sequences for the corresponding mRNAs. A previously recognized but unidentified long and relatively rare cDNA clone containing the entire sequence from the cap site at the 5' end to the poly(A) tail at the 3' end of the mRNA was isolated by single-step screening of the library. The size, amino acid composition, and the activity of the protein expressed in heterologous cells strongly suggest this mRNA codes for lysophospholipase [Van den Bosch, H., Aarsman, A. J., DeJong, G. N., & Van Deenen, L. M. (1973) Biochim. Biophys. Acta 296, 94-104].

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cattle
  • Cloning, Molecular*
  • DNA / isolation & purification*
  • Genetic Vectors
  • Lysophospholipase / genetics*
  • Pancreas / enzymology*
  • Phospholipases / genetics*
  • RNA, Messenger / genetics*
  • RNA, Messenger / isolation & purification
  • Rats
  • Species Specificity

Substances

  • RNA, Messenger
  • DNA
  • Phospholipases
  • Lysophospholipase

Associated data

  • GENBANK/M15893