Measurement of human prolactin messenger RNA in decidual tissues using complementary DNA probe cloned in M13mp9 bacteriophage.
A human prolactin (PRL) cDNA clone was digested with restriction enzyme Pst I and the resultant fragments were cloned into bacteriophage M13mp9. Single stranded recombinant DNAs having a coding strand of the PRL cDNA were selected by hybridization with 125I-labeled PRL mRNA obtained from human prolactinoma tissue. One of the single stranded recombinant DNAs was purified by agarose gel electrophoresis and labeled with 125I to a specific activity of 1.4 X 10(8) cpm per microgram of DNA. The probe could be successfully used in RNA dot hybridization. Analysis of poly (A) RNAs from prolactinoma, liver and placental tissues revealed that this probe was specific to PRL mRNA sequence. Hybridization of poly (A) RNA from decidual tissue to this probe revealed the presence of PRL mRNA sequence. However, PRL mRNA in decidua was at least 20,000 times less than that in pituitary prolactinoma.
D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D003656:Decidua / Q000032:analysis*; D004926:Escherichia coli; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization; D010911:Pituitary Neoplasms / Q000032:analysis / Q000378:metabolism; D011247:Pregnancy; D011388:Prolactin / Q000032:analysis* / Q000378:metabolism; D012317:RNA Phages; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger; D011388:Prolactin; D004262:DNA Restriction Enzymes