YU Xiao-long, TANG Li-jie and LI Yi-jing. Study on Inhibition of TGEV Replication by Antisense RNA[J]. Virologica Sinica, 2004, 19(3): 268-270.
Citation: YU Xiao-long, TANG Li-jie, LI Yi-jing. Study on Inhibition of TGEV Replication by Antisense RNA .VIROLOGICA SINICA, 2004, 19(3) : 268-270.

反义RNA抑制猪传染性胃肠炎病毒复制的研究

  • 通讯作者: 李一经*, 
  • 根据反义RNA作用原理,设计一条互补猪传染性胃肠炎病毒基因(26888-27184)区的反义RNA序列。将该序列与逆转录病毒表达载体构建成质粒PLXSN-N5’,并与质脂体共转染PA317细胞,经G418(500 μg/ml)筛选出稳定的产毒细胞克隆。取其上清液感染小鼠成纤维细胞NIH3T3,测定细胞克隆产生的假病毒滴度,用高滴度假病毒感染IBRS2细胞。提取被感染的IBRS2细胞总DNA和RNA,通过PCR和RT-PCR证明PLXSN-N5’整合到IBRS2细胞基因组。病毒感染细胞病变表明,反义RNA有明显抑制TGEV复制的作用。

Study on Inhibition of TGEV Replication by Antisense RNA

  • Corresponding author: LI Yi-jing, 
  • In the study, we designed an antisense RNA targeting at the highly conserved sequence (26880-27184nt)of Transmissible gastroenteritis virus of swine,(TGEV). The fragment cloned by PCR was recombined with retrovirus expression vector, pLXSN, and transfected into packaging cell line PA317 with Lipofectamine. The viral supenatants of the clones selected with G418 (500 g/mL) were detected by murine fibroblast NIH3T3. The highest viral titer pseudovirus was used to infect IBRS2 cell. PCR and RT-PCR analysis indicated that the pLXSN-N5’ had inserted into the genome of IBRS2 cell. The examination of cytopathic effect (CPE) showed that the antisense RNA could inhibit TGEV replication effectively.

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    Study on Inhibition of TGEV Replication by Antisense RNA

      Corresponding author: LI Yi-jing,
    • 1. Animal medicine College of Northeast Agricultural University, Harbin 150030, China

    Abstract: In the study, we designed an antisense RNA targeting at the highly conserved sequence (26880-27184nt)of Transmissible gastroenteritis virus of swine,(TGEV). The fragment cloned by PCR was recombined with retrovirus expression vector, pLXSN, and transfected into packaging cell line PA317 with Lipofectamine. The viral supenatants of the clones selected with G418 (500 g/mL) were detected by murine fibroblast NIH3T3. The highest viral titer pseudovirus was used to infect IBRS2 cell. PCR and RT-PCR analysis indicated that the pLXSN-N5’ had inserted into the genome of IBRS2 cell. The examination of cytopathic effect (CPE) showed that the antisense RNA could inhibit TGEV replication effectively.

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