Cloning and chromosomal localization of a gene encoding a novel serine/threonine kinase belonging to the subfamily of testis-specific kinases

Pablo E. Visconti, Zhonglin Hao, Marie A. Purdon, Paula Stein, Binaifer R. Balsara, Joseph R. Testa, John C. Herr, Stuart B. Moss, Gregory S. Kopf

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

Using reverse transcription-polymerase chain reaction (RT-PCR) with degenerate oligonucleotides corresponding to two highly conserved motifs within the protein kinase family of catalytic domains, we isolated a PCR fragment encoding a novel member of the testis-specific serine/threonine kinases (STK) from mouse male mixed germ cell mRNA. This PCR fragment recognized a 1020-bp transcript in male germ cells by northern blot analysis and was used to clone a full-length cDNA from a mouse mixed germ cell cDNA library. This cDNA has an open reading frame of 804 bases encoding a protein of 268 amino acids. This novel gene is almost identical to Stk22c, encoding a recently described testis-specific protein kinase, except for base-pair deletions that result in a shift in the coding region and an alteration of 22 amino acids (residues 109-131). Due to its homology with Stk22c, we have called this protein kinase gene Stk22d. Northern blot analysis revealed that this protein kinase is developmentally expressed in testicular germ cells and is not present in brain, ovary, kidney, liver, or early embryonic cells. We then cloned the human homologue of this protein kinase gene (STK22C) and found it to be expressed exclusively in the testis. Fluorescence in situ hybridization with both the human and mouse cDNA clones revealed syntenic localization on chromosomes 1p34-p35 and 4E1, respectively.

Original languageEnglish
Pages (from-to)163-170
Number of pages8
JournalGenomics
Volume77
Issue number3
DOIs
StatePublished - Aug 15 2001

Keywords

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Chromosome Mapping
  • Chromosomes, Human, Pair 1/genetics
  • Cloning, Molecular
  • DNA, Complementary
  • Female
  • Gene Expression
  • Gene Library
  • Humans
  • In Situ Hybridization, Fluorescence
  • Male
  • Mice
  • Molecular Sequence Data
  • Organ Specificity
  • Protein Serine-Threonine Kinases/chemistry
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Homology, Nucleic Acid
  • Spermatozoa/enzymology
  • Testis/enzymology

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