Cloning and expression of cynomolgus monkey (Macaca fascicularis) gonadotropins luteinizing hormone and follicle-stimulating hormone and identification of two polymorphic sites in the luteinizing hormone beta subunit

The genes encoding the cynomolgus monkey gonadotropin subunits, alpha, foll icle-stimulating hormone (FSH) beta and luteinizing hormone (LH) beta, were cloned by reverse transcriptase polymerase chain reaction (RT-PCR) using p ituitary RNA. The predicted amino acid sequences displayed 82, 96 and 87% i dentity to human subunit sequences, respectively. Northern blot hybridizati on of monkey tissues revealed pituitary specific transcripts of 1.0 and 0.6 kb for the alpha and LH beta subunit, respectively, and two bands of 1.8 a nd 0.65 kb for the FSH beta. Upon sequencing LH beta cDNAs from different m onkeys, two polymorphic sites were detected, resulting in the amino acid tr ansitions Ser32Thr and His60Arg. Restriction analysis revealed different ho mo- and heterozygous combinations of the polymorphic sites indicating linka ge dysequilibrium. Transient co-expression of the alpha subunit together wi th the FSH beta or LH beta subunit in COS7 and CHO cells resulted in secret ion of in vitro bioactive hormones. This work represents a further step tow ards production of recombinant monkey LH and FSH which can be used in a hom ologous experimental setting in the cynomolgus monkey. (C) 1999 Elsevier Sc ience Ireland Ltd. All rights reserved.