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

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].

[1]  S. Rogers,et al.  Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. , 1986, Science.

[2]  W. Rutter,et al.  Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose , 1986, Cell.

[3]  U. Krawinkel,et al.  Rapid synthesis of cDNA for cloning into lambda vectors. , 1986, Nucleic acids research.

[4]  E. Chen,et al.  Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. , 1985, DNA.

[5]  W. Rutter,et al.  Isolation and sequence of a rat chymotrypsin B gene. , 1984, The Journal of biological chemistry.

[6]  H. Kern,et al.  Hormonal stimulation in the exocrine pancreas results in coordinate and anticoordinate regulation of protein synthesis , 1984, The Journal of cell biology.

[7]  M E Watson,et al.  Compilation of published signal sequences. , 1984, Nucleic acids research.

[8]  M. Kozak.  Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. , 1984, Nucleic acids research.

[9]  R. W. Davis,et al.  Yeast RNA polymerase II genes: isolation with antibody probes. , 1983, Science.

[10]  R. W. Davis,et al.  Efficient isolation of genes by using antibody probes. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[11]  B. Howard,et al.  Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells , 1982, Molecular and cellular biology.

[12]  Raymond J MacDonald,et al.  Two similar but nonallelic rat pancreatic trypsinogens. Nucleotide sequences of the cloned cDNAs. , 1982, The Journal of biological chemistry.

[13]  W. Rutter,et al.  Primary structure of two distinct rat pancreatic preproelastases determined by sequence analysis of the complete cloned messenger ribonucleic acid sequences. , 1982, Biochemistry.

[14]  H. Okayama,et al.  High-efficiency cloning of full-length cDNA , 1982, Molecular and cellular biology.

[15]  H. Hauser,et al.  5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency. , 1981, Nucleic acids research.

[16]  W. Rutter,et al.  Structure of a family of rat amylase genes , 1980, Nature.

[17]  W. Rutter,et al.  Proteins synthesized and secreted during rat pancreatic development , 1980, The Journal of cell biology.

[18]  W. Rutter,et al.  Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. , 1979, Biochemistry.

[19]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[20]  J. Depierre.  Extraction and assay of small quantities of free fatty acids produced by hydrolysis of microsomal membranes. , 1977, Analytical biochemistry.

[21]  W. Rutter,et al.  Rat insulin genes: construction of plasmids containing the coding sequences. , 1977, Science.

[22]  H. Pelham,et al.  An efficient mRNA-dependent translation system from reticulocyte lysates. , 1976, European journal of biochemistry.

[23]  T. Rabbitts,et al.  Bacterial cloning of plasmids carrying copies of rabbit globin messenger RNA , 1976, Nature.

[24]  F. Stahl,et al.  Rec-mediated recombinational hot spot activity in bacteriophage lambda. III. Chi mutations are site-mutations stimulating rec-mediated recombination. , 1975, Journal of molecular biology.

[25]  A. van der Eb,et al.  A new technique for the assay of infectivity of human adenovirus 5 DNA. , 1973, Virology.

[26]  J. D. de Jong,et al.  Studies on lysophospholipases. I. Purification and some properties of a lysophospholipase from beef pancreas. , 1973, Biochimica et biophysica acta.

[27]  P. Leder,et al.  Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[28]  W. Rutter,et al.  Selective expression of rat pancreatic genes during embryonic development. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[29]  C. Coleclough,et al.  Use of primer-restriction-end adapters in a novel cDNA cloning strategy. , 1985, Gene.

[30]  E. Chen,et al.  Supercoil sequencing: a fast method for sequencing plasmid DNA , 1985 .

[31]  J. Martial,et al.  A method for isolation of intact, translationally active ribonucleic acid. , 1983, DNA.

[32]  W. Rutter,et al.  Rat preprocarboxypeptidase A: cDNA sequence and preliminary characterization of the gene. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[33]  W. Gilbert,et al.  Sequencing end-labeled DNA with base-specific chemical cleavages. , 1980, Methods in enzymology.