Tilapia prolactin: molecular cloning of two cDNAs and expression in Escherichia coli.
We have isolated cDNA clones encoding tilapia prolactin (tiPRL) from a cDNA library prepared from tilapia (Oreochromis niloticus) anterior pituitary glands. A trout PRL cDNA fragment was used as hybridization probe to select the recombinant plasmids carrying the tiPRL coding sequence. Two types of PRL cDNA were isolated and their complete nucleotide sequence determined. The larger cDNA (tiPRL-I) codes for a polypeptide of 212 amino acids, including a putative signal sequence of 24 amino acids, and contains a 3' untranslated region of 787 bp. The second prolactin cDNA (tiPRL-II) encodes a polypeptide of 200 amino acids, including a presumptive signal peptide of 23 amino acids, and contains a noncoding region of 512 bp. tiPRL-I and tiPRL-II cDNA sequences are 81% similar, whereas the encoded proteins share 69% amino acid identity at optimal alignment. Mature tiPRL-I was efficiently expressed in Escherichia coli carrying a plasmid in which the tiPRL-I cDNA was under the control of the phi 10 promoter of T7 bacteriophage. The new recombinant protein representing about 45% of the total cellular proteins was found in inclusion bodies and cross-reacted with salmon PRL antiserum.
Rentier-Delrue
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F
Laboratoire Central de Génie Génétique, Université de Liège, Belgium.
Swennen
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D
Prunet
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Lion
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Martial
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D005399:Fishes / Q000235:genetics; D006801:Humans; D008322:Mammals / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011388:Prolactin / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics; D015183:Restriction Mapping; D012487:Salmonidae / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014337:Trout / Q000235:genetics*
D021382:Protein Sorting Signals; D011388:Prolactin; D004247:DNA