Molecular cloning and expression of salmon prolactin cDNA.
Prolactin was purified from chum salmon pituitaries. It was resolved into two variants by reverse-phase high-performance liquid chromatography. A cDNA library was prepared from Pacific chinook salmon pituitaries. Salmon prolactin gene was screened using a synthetic oligonucleotide based on partial protein sequence. A positive clone (PRL-10) was identified and sequenced. It is a full-size clone containing 1.1 kb and coding for a preprolactin of 211 amino acids. A modified prolactin plasmid (PRL-10A), in which the 5' untranslated sequence and the nucleotide sequence coding for the signal peptide of prolactin were deleted, was reconstructed into an expression vector using the heat-inducible lambda pL promotor. Mature prolactin, a single polypeptide of 22 kDa, was efficiently expressed in the bacteria at an elevated temperature.
Song
S
S
Research Institute, Hospital for Sick Children, Toronto, Canada.
Trinh
K Y
KY
Hew
C L
CL
Hwang
S J
SJ
Belkhode
S
S
Idler
D R
DR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis; D010902:Pituitary Gland / Q000032:analysis; D010957:Plasmids; D011388:Prolactin / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D012474:Salmon / Q000235:genetics; D012756:Sheep; D013045:Species Specificity
D010455:Peptides; D012333:RNA, Messenger; D011388:Prolactin; D004247:DNA