【摘要】 以转基因处理的家蝇卵为对象,采用银染差异显示技术(DD-PCR),对携带绿色荧光蛋白(GFP)和抗菌肽(ABP)融合基因的真核表达质粒以电转移的方式进入家蝇受精卵后,卵内基因的表达情况做了初步研究。结果表明,转基因处理的蝇卵与未经转基因处理的蝇卵在分化开始之前基因的表达存在差异;以T11MA为锚定引物,在转基因处理组发现6条差异表达的片段,成功地克隆得到其中3条。序列分析证实其中2条为新基因片段,另1条为家蝇细胞色素C的编码片段;斑点杂交证实了新基因片段仅在转基因处理组得到表达,而细胞色素C在2组中都有表达。获得的新基因片段登录于GenBank,登录号分别为AY030235和AY030236。本试验结果为解释外源基因在宿主细胞内的去向,发现宿主内部对目的基因整合和分解的相关因素提供了可能,并为系统地开展发育相关研究打下了基础。更多还原

【Abstract】 The transcriptional process of the early housefly embryos processed by transgene was studied and compared with the non-processed embryos by differential display-PCR(DD-PCR).The housefly embryos were electroporated with eukaryotic expression plasmids——pcmGS1 containing GFP(green fluorescent protein) and ABP (antibacterial peptides) fusion gene.The situation of gene expression was studied by DD before differentiation of the embryos,and the differences between the normal and processed group were found.Three differential fragments of transgenic groups were cloned.The results of sequencing and homology analysis showed that one of them was a part of cytochrome C gene of Musca domestica,and the others 240 nt and 432 nt were new sequences.Thirdly,significant signals were observed when the two new fragments(genes) were hybridized with housefly genome by dot-blot.The results indicated that they may be the part of housefly genome.Their squences had been signed in the GenBank with their registered codons to be of AY030235 and AY030236,respectively.The fragment of cytochrome C gene was confirmed by Northern blot.The result showed that the gene was expressed in both testing and normal embryos,which would be a quantity difference:the cDNA quantity of cytochrome C expressed in testing group was more than in normal group.更多还原