Development of the virus-control system in pigs using the ribozyme

核糖酶在猪病毒控制系统中的应用核糖酶能够催化RNA的序列特异性裂解。因此，它们已成为通过切割rna人工控制基因表达的潜在工具。为选育抗伪狂犬病毒（PRV）引起的Aujeszkey病猪，设计了该病毒直接早180基因mRNA的发夹核酶，并利用猪培养细胞检测其活性。将转录RNA聚合酶II启动子（CMV增强子和启动子）或RNA聚合酶III启动子（tRNA）的核糖酶基因引入猪培养细胞。用RNA聚合酶II转录核酶基因的培养细胞感染PRV后，在感染后第1天观察到CPE。然后CPE在三天内扩散到整个培养液。这一结果与未引入核酶基因的猪培养细胞的结果基本相同。然而，RNA聚合酶III转录核酶基因的培养细胞显示出对PRV感染的抗性，感染后6天CPE没有在整个培养物中传播。

Development of the virus-control system in pigs using the ribozymeRibozymes are capable of catalyzing sequence specific cleavage of RNA.Therefore they have been emerging as potential tools to artificially control gene expression through cleavage of RNAs. For breeding of the pig which has resistance to Aujeszkey's disease caused by the infection of pseudorabies virus (PRV), we designed hairpin ribozyme for mRNA of immediate early 180 gene of the virus and examined the activity of ribozyme using pig cultured cell. The riobzyme gene transcribed the promoter of RNA polymerase II (CMV enhancer and promoter) or III (tRNA) was introduced to the pig cultured cells. The cultured cell, in which the ribozyme gene was transcribed by RNA polymerase II was infected with PRV, CPE was observed in cultured cell on the first day after infection. And then CPE spread over the whole culture within three days. This result was almost the same as that in the pig cultured cell which ribozyme gene was not introduced. However, the cultured cell, in which the ribozyme gene was transcribed by RNA polymerase III, revealed the resistance to PRV infection showing that CPE didn't spread over the whloe culture by six days after infection.