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Cloning and expression analysis of putative glyceraldehyde-3-phosphate dehydrogenase genes in Pilobolus crystallinus

机译:毛状脑毛虫推定的3-磷酸甘油醛脱氢酶基因的克隆与表达分析

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Putative glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes were cloned from Pilobolus crystallinus with degenerated primers designed from conserved sequences in many GAPDHs. P. crystallinus had three gapdh homologue genes, named pcgapdh1, pcgapdh2, and pcgapdh3. Deduced amino acid sequences for PCGAPDH1, PCGAPDH2, and PCGAPDH3 showed highest similarity with GPD3, GPD1, and GPD2, respectively, of Mucor circinelloides, indicating that these three gapdh genes had diverged before Pilobolus and Mucor were separated. The expression patterns of the gapdh genes, however, were quite different between P. crystallinus and M. circinelloides. All the three pcgapdh genes were expressed, and the expression of pcgapdh2 was suppressed by glucose and sodium acetate. These results indicate that the function of these orthologous genes was changed after Pilobolus and Mucor were separated.
机译:利用从许多GAPDHs中的保守序列设计的简并引物,从毛状毛果克隆了推定的3-磷酸甘油醛脱氢酶(gapdh)基因。 P. crystallinus具有三个gapdh同源基因,分别名为pcgapdh1,pcgapdh2和pcgapdh3。推导的PCGAPDH1,PCGAPDH2和PCGAPDH3的氨基酸序列分别与Mucor circinelloides的GPD3,GPD1和GPD2具有最高的相似性,表明这3个gapdh基因在发毛ilo虫和Mucor分离之前就已经发散了。然而,gapdh基因的表达模式在结晶青霉和圆环青霉之间是完全不同的。表达了全部三个pcgapdhdh基因,并且葡萄糖和乙酸钠抑制了pcgapdh2的表达。这些结果表明,在分离毛毛虫和Mucor后,这些直系同源基因的功能发生了改变。

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