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Endocrinology, Vol 133, 1744-1752, Copyright © 1993 by Endocrine Society
ARTICLES |
TG Golos, M Durning, JM Fisher and PD Fowler
Wisconsin Regional Primate Research Center, University of Wisconsin, Madison 53715-1299.
The close homology of the chorionic somatomammotropin (CS) genes with pituitary GH is unique to primates. Southern blots of rhesus genomic DNA probed with a human CS cDNA demonstrated that the rhesus gene family consists of at least five EcoRI fragments between 2.3-9.5 kilobases. Rhesus pituitary and placental cDNA libraries were screened for hCS-hybridizing clones. A pituitary GH cDNA was isolated that was 95% and 96% identical to human GH at the mRNA and protein levels, respectively. Thirteen placental clones representing four different cDNAs were identified and sequenced. The relationships of the rhesus placental clones among themselves were similar to those among human placental mRNAs. Two cDNAs (mCS1 and mCS2) differed by two bases and coded for identical proteins. A third cDNA (mCS3) was 94% homologous to mCS1/2 in mRNA sequence as well as in deduced amino acid sequence. A fourth cDNA (mGH-V) was 84-86% homologous to the other placental cDNAs at the mRNA and protein levels, similar to the homology between human placental GH-V and CS mRNAs. To determine the relative expression of placental mRNAs, we developed a quantitative reverse transcription- polymerase chain reaction assay using diagnostic NlaIV restriction enzyme sites in cDNAs amplified from the placental mRNAs. All four mRNAs were expressed in the placenta throughout most of pregnancy, with the mCS1 and mGH-V mRNAs generally expressed at 2- to 10-fold higher levels than mCS2 and mCS3. Additionally, mCS3 and mGHV were expressed at relatively higher levels during the second and third trimesters than during the first trimester.
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