The Application of PCR-DGGE Technique in Sheeplntestinai Microbial Diversity Research

摘要

To identify the intestinalmicrofloradiversityoftransgenic sheep and non-transgenic one with overexpression of foreign antiviral gene (VP1 & SB-SW),DNA of total bacteriawereextracted from 16 sheep fecal samples(4 non-transgenic sheep, 6 VP1 transgenic sheep and 6 SB-SW transgenic sheep). And then PCR amplification with bacterial universalprimers of V3 variable region of 16S rRNA weretaken to get the fingerprint profile bydenaturing gradient gel electrophoresis (DGGE) technology. The results showed thatthe DGGE profiles of the 16fecal samples were highly polymorphic.The number of DGGE bands, considered indicative of the total species richness, did not vary predictably among the threedifferent samples.The DNAsequences were analyzed and the dominant bacteria in sheep fecal were found to beBacteroides, Clostridium, and Ruminococ-cus.Specially,non-transgenic sheep had more Alistipesfinegoldii and Clostridiumlentocellum, transgenic sheep with VP1 hadmore Clostridiumdrakei and Clostridium populeti and transgenic sheep with SB-SW had moreBarnesiellaintestinihominis and Clostridiumljungdahlii. So the PCR-DGGE technique can be applied toevaluategenetically modified (GM) animal potential risks to the environment.