The N-terminal half of talin2 is sufficient for mouse development and survival

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Abstract

Using a talin2 gene-trapped embryonic stem cell clone, we have developed a talin2 mutant mouse line that expresses the N-terminal half (1–1295) of talin2 fused with β-galactosidase. The homozygous mutant mice appear to be normal and healthy. In the testis, talin2 expresses as a shorter form with a unique 30 residues at N-terminus linking to a common C-terminus from 1122 to 2453 of the long form. The resulting talin2 in the mutant testis only contains 204 residues of the wild-type testis talin2. However, it did not seem to affect the morphology of testis or reproduction of male mice. In fact, male and female mutant mice are fertile. Utilizing the expression of talin2(1–1295)/β-galactosidase fusion protein, we have examined the distribution of talin2 in tissues. In contrast to talin1, talin2 expression is more restricted in tissues and cell types.

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Materials and methods

Embryonic stem (ES) cells and mice. Talin2in1295 gene-trap SV129 ES cells (RRI434) were obtained from BayGenomics (UCSF) and injected into the C57BL/6 blastocysts. The resulting male chimeras were mated with C57BL/6 females, and the talin2in1295 line was subsequently maintained on a mixed C57BL/6:SV129 genetic background. Breeding was carried out under standard animal housing conditions in the UCD animal facility.

Genotyping was performed by polymerase chain reaction (PCR) assay. Mouse tail

Interruption of talin2 gene in mice

We have searched the available databases for mouse talin2 gene and two partial cDNA sequences (GenBank Accession Nos. AF467081 and BC059856) were identified. Together with the talin2 sequence from mouse chromosome 9 (XP_486227), we have assembled the mouse talin2 cDNA encoding 2453 amino acid residues, which shows 73% identity (85% similarity) with mouse talin1 (2541 amino acids, NM_011602) (Fig. 1).

By searching the BayGenomics database, we have found three gene-trapped embryonic stem (ES) cell

Discussion

In addition to the mouse, the existence of a second talin gene was reported in humans [12] and Dictyostelium[16]. We also found partial sequences of talin2 in chimpanzee (Pan troglodytes), rat (Rattus norvegicus), cow (Bos taurus), dog (Canis familiaris), red jungle fowl (Gallus gallus), and zebrafish (Danio rerio) from EST databases. Their derived amino acid sequences are extremely conserved. For example, human (NM_015059), rat (XP_236367), and mouse are 95% identical. However, the mammalian

Acknowledgments

We thank the BayGenomics for providing the gene-trapped ES clone. This study is supported in part by Shriner’s Children Hospital grant (S.H.L.).

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