ABSTRACT
<p><b>OBJECTIVE</b>To construct the eukaryotic expression plasmid containing lvgA gene flanked with CpG motifs of Legionella pneumophila for its expression in NIH3T3 cells.</p><p><b>METHODS</b>lvgA gene flanked with CpG motifs of Legionella pneumophila was amplified by PCR. The PCR products was inserted into the eukaryotic expression plasmid pcDNA3.1/myc-his(+) to construct the recombinant plasmid pclvgA/CpG, which was subsequently transfected into NIH3T3 cells via lipofection. Immunofluorescence analysis was carried out to detect the transient expression of the plasmid in the cells.</p><p><b>RESULTS</b>Sequence analysis showed that the recombinant plasmid pclvgA/CpG contained the lvgA/CpG fragment with a length of 657 bp, encoding a protein of 27.7 Ku. Immunofluorescence analysis identified the transient expression of the recombinant plasmid pclvgA/CpG in NIH3T3 cells.</p><p><b>CONCLUSION</b>The lvgA gene flanked with CpG motifs of Legionella pneumophila has been constructed successfully, and the transient expression of the recombinant plasmid pclvgA/CpG can be detected in NIH3T3 cells.</p>
Subject(s)
Animals , Mice , Bacterial Vaccines , Genetics , CpG Islands , Genetics , Legionella pneumophila , Genetics , NIH 3T3 Cells , Plasmids , Genetics , Recombinant Proteins , Genetics , Allergy and Immunology , Transfection , Virulence Factors , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the characteristics and distribution of GATA-4 in the testis of male mice.</p><p><b>METHODS</b>Paraffin sections were obtained from the testes of 24 male B6SJLF1/J mice, aged 0 day (n = 6), 2 weeks (n = 6), 4 weeks (n = 6) and 6 weeks (n = 6), and the expressions of GATA-4 in the testis were observed by the immunohistochemical ABC method and DAB visualization at different times.</p><p><b>RESULTS</b>Positive expressions of GATA4 were found in the Sertoli cells and Leydig cells of all the mice, but significantly higher in the 4- and 6-week-old than in the 0-day and 2-week-old groups (P < 0.01). And they were also observed in the germ cells of the 4- and 6-week-old mice, significantly higher in the latter than in the former (P < 0.01).</p><p><b>CONCLUSION</b>GATA-4 exists in the testis of male mice, which has provided a morphological base for sex determination and differentiation and hormone regulation in the testis.</p>
Subject(s)
Animals , Male , Mice , Cell Differentiation , GATA4 Transcription Factor , Metabolism , Germ Cells , Metabolism , Leydig Cells , Metabolism , Sertoli Cells , Metabolism , Testis , Cell Biology , MetabolismABSTRACT
The GATA family proteins are a group of zinc finger transcription factors that are expressed in human and mammalian animals and play an important role in mammalian organ morphogenesis, cell proliferation and sex differentiation. GATA-4 and GATA-6 have been identified in the ovaries and testes of humans, mice, pigs and chickens. GATA-4 contributes to fetal male gonadal development by regulating the genes that mediate Müllerian duct regression and the onset of testosterone production. GATA-4 and GATA-6 are localized in and regulate the function of the ovarian and testicular somatic cells of fetal mice, especially granulosa cells, thecal cells, Sertoli cells and Leydig cells. GATA-4 is also present in the germ cells of fetal and prepubertal mice.