中文摘要：

To explore the pluripotency maintenance and update the functional influence of pluripotency genes c Nanog and c Pou V in chicken( Gallus gallus) embryonic stem cells( c ESCs),the stable RNAi vectors p Super-c Nanog and p Super-c Pou V constructed previously were used to transfect c ESCs. The mRNA levels of two target genes were detected with real-time PCR. These two genes were down-regulated since the 48 thh and the down-regulation continued with the extension of time,the interference efficiency reached 65% at 96thhour( P < 0. 05). With the down-regulation of c Nanog or c Pou V gene,c ESCs showed differentiation and proliferation rate of these cells slowed down,the domed colony of these cells disappeared gradually when the edge of colony became irregular. At 96 thhour after transfection,the alkline phosphatase( AKP) and stage-specific embryonic antigen-1( SSEA-1) were not be detected in c Nanog gene-knocked out c ESCs,but it was done in that with c Pou V gene-knocked out. The c Pou V-suppressing c ESCs were again transfected with p Super-c Nanog,the pluripotency markers AKP and SSEA-1 were both not found expressing at the 48 thhour. The results showed that c Pou V and c Nanog genes played an important role in maintaining pluripotency and self- renewal in c ESCs,and c Nanog gene was dominant. To sum up,our results may provide insights into the molecular regulation mechanism of avian during development.

英文摘要：

To explore the pluripotency maintenance and update the functional influence of pluripotency genes cNanog and cPouV in chicken （ C,a/lus gallus） embry- onic stem cells （ cESCs）, the stable RNAi vectors pSuper-cNanog and pSuper-cPouV constructed previously were used to transfect cESCs. The mRNA levels of two target genes were detected with real- time PCR. These two genes were down-regulated since the 48^th and the down-reg-lation continued with the extension of time, the interference efficiency reached 65% at 96^th hour （P 〈0.05）. With the down-regulation of cNanog or cPouV gene, cESCs showed differentiation and prolifera- tion rate of these cells slowed down, the domed colony of these cells disappeared gradually when the edge of colony became irregular. At 96^th hour after transfection, the alkline phosphatase （AKP） and stage-specific embryonic antigen-1 （ SSEA-1 ） were not be detected in cNanog gene-knecked out eESCs, but it was done in that with cPouV gene -knocked out. The cPouV-suppressing cESCs were again transfected with pSuper-cNanog, the pluripotency markers AKP and SSEA-1 were both not found expressing at the 48^th hour. The results showed that cPouV and cNartog genes played an important role in maintaining pluripotency and self- renewal in cESCs, and cNanog gene was dominant. To sum up, our results may provide insights into the molecular regulation mechanism of avian during development.