PMID: 7543295Jun 1, 1995Paper

Partial complementary deoxyribonucleic acid cloning of equine relaxin messenger ribonucleic acid, and its localization within the equine placenta

Biology of Reproduction
T KlonischD G Porter

Abstract

To determine the site of relaxin gene expression in equine placentae, a set of degenerate oligonucleotide primers was made according to the published amino acid sequence of the A- and B-chain of equine relaxin (eRXN). Total cellular RNA (tcRNA) from equine placentae at about 120 and 300 days of pregnancy was subjected to reverse transcriptase-polymerase chain reaction (RT-PCR) with use of these primers. A single amplification product of approximately 430 bp was detected in each case by agarose gel electrophoresis. The PCR product was ligated into Bluescript plasmid and sequenced to confirm the identity of the clone as an eRXN cDNA fragment. Nucleic acid sequence analysis revealed a 428-bp eRXN cDNA fragment encoding for parts of the A- and B-chain and the connecting peptide (109 residues). Northern analysis of tcRNA from placentae of 120 and 300 days of pregnancy was carried out with use of antisense digoxigenin-labeled cRNA generated from the cDNA clone, and a single transcript of approximately 1 kb was detected. In situ hybridization on placental tissue at 120 and uteroplacental tissue at 300 days of pregnancy indicated that only the fetal trophoblastic cells expressed eRXN mRNA transcripts. The identity of these cells was co...Continue Reading

Citations

Feb 25, 2000·Molecular and Cellular Endocrinology·S Hombach-KlonischT Klonisch
Mar 1, 1997·Placenta·T KlonischW R Allen
Feb 20, 1999·Biology of Reproduction·T KlonischB Fischer
Mar 22, 2000·Biology of Reproduction·S Hombach-KlonischT Klonisch
Mar 7, 2001·Molecular Biology and Evolution·T KlonischS Hombach-Klonisch

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