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目的利用基因芯片技术筛选与精子发生相关的基因,并研究其表达特点。方法将4d、9d、18d、35d、54d龄和6月龄小鼠睾丸组织cDNA探针与Affymetrix全基因组芯片进行杂交,筛选出差异表达的基因。然后通过RT-PCR分析差异表达基因在小鼠睾丸不同发育阶段中的表达。结果对Affymetrix全基因组芯片杂交结果分析后,筛选得到1个差异表达杂交点,通过在NCBI网站与小鼠全基因组序列Blast分析可知该差异表达基因是Pgk2基因。该基因在4d、9d、1 8d、35d、54d龄和6月龄小鼠睾丸中信号强度分别是1.7(A)、161.5(P)、634(P)、1881.3(P)、3123.7(P)和3440.4(P)表明Pgk2基因在4d龄小鼠睾丸中无表达,9d低表达,从18年龄后开始高表达;RT-PCR结果表明:小鼠Pgk2基因在小鼠9d龄及之前的睾丸中无表达或低表达,在18d龄睾丸后开始高表达,与芯片数据的结果相一致。结论 Pgk2基因为小鼠年龄依赖性表达基因,推测其在精子发生过程中有重要作用。
Objective To screen the genes related to spermatogenesis by gene chip technology and study its expression characteristics. Methods The cDNA probes of testis in 4d, 9d, 18d, 35d, 54d and 6-month-old mice were hybridized with Affymetrix genome-wide microarray to screen differentially expressed genes. Then RT-PCR analysis of differentially expressed genes in mouse testes at different stages of development. Results After analysis of Affymetrix genome-wide hybridization results, one differentially expressed hybridization point was screened. Blast analysis showed that the differentially expressed gene was Pgk2 gene on NCBI website and mouse genome-wide. The signal intensity of the gene in testis of mice at 4d, 9d, 18d, 35d, 54d and 6w were 1.7 A, 161.5 P, 634 P, 1881.3 P, 3123.7 P, And 3440.4 (P) showed that the Pgk2 gene was not expressed in testis of 4-d old mice and was low in 9d, and was overexpressed after 18-year-old. The results of RT-PCR showed that the Pgk2 gene in mouse testis No expression or low expression, high expression began after 18 days of testes, consistent with the results of the chip data. Conclusion The Pgk2 gene is an age-dependent gene expressed in mice, suggesting that it plays an important role in the process of spermatogenesis.