The gene for murine megakaryocyte growth and development factor (thrombopoietin, Thpo) is located on mouse chromosome 16

Abstract

The canine MGDF genomic clone was isolated using a degenerative oligonucleotide probe based on the amino terminal sequences of the MGDF protein purified from aplastic canine serum. A mouse genomic library (1 x 10{sup 6} pfu) was then screened with a mixture of six canine MGDF oligomers. Phage DNA was isolated from the positive clone and sequenced. Comparison of the murine MGDF and the human MGDF gene showed that their genomic structures are similar in sizes of exons and introns. To determine chromosomal location of the murine MGDF gene (locus designation thrombopoietin, Thpo), interspecific backcross progeny were generated by mating (C57BL/6J X Mus spretus) F{sub 1} females and C57BL/6J males as described. This interspecific backcross mapping panel has been typed for over 1700 loci that are well distributed among all of the autosomes as well as the X chromosome. A total of 205 N{sub 2} mice were used to map the Thpo locus. The probe used in these studies was an {approximately}440-bp fragment of mouse genomic DNA consisting of exon 6, intron 6, and part of exon 7. C57BL/6J and M. spretus DNAs were digested with several enzymes and analyzed by Southern blot hybridization for informative restriction fragment length polymorphismsmore » (RFLPs) using a mouse genomic probe. A fragment of 6.5 kb was detected in EcoRI-digested C57BL/6J DNA, and a fragment of 4.6 kb was detected in EcoRI-digested M. spretus DNA. The 4.6-kb M. spretus-specific EcoRI fragment was used to follow the segregation of the Thpo locus in backcross mice. 5 refs., 1 fig.« less