Fang Zheng, Shao Jingxu, Weng Qingbei
School of Life Sciences, Guizhou Normal University, Guiyang, 550001, China.
Virol Sin. 2016 Oct;31(5):425-436. doi: 10.1007/s12250-016-3791-8. Epub 2016 Oct 18.
Cells of the P8-Se301-C1 strain are Spodoptera exigua cell clones that each harbor a partial version of the S. exigua multiple nucleopolyhedrovirus (SeMNPV) genome and which are resistant to homologous SeMNPV infections. The cells produce no viral progeny, suggesting that the infection is a latent-like viral infection. To investigate the SeMNPV genes harbored in the P8-Se301-C1 cells, the de novo transcriptomes of P8-Se301-C1 cells and S. exigua Se301 cells were analyzed and compared. A total of 54,569,296 reads were obtained from the P8-Se301-C1 cells that yielded 112,565 final unigenes with a mean length of 1,093 nt. A total of 56,865,504 reads were obtained from the Se301 cells that yielded 102,996 final unigenes with a mean length of 1,082 nt. Ten SeMNPV gene transcripts (se5, se7, se8, se12, se43, se45, se89, se90, se124, and se126) were detected in the P8-Se301-C1 cells by RNA-Seq but not in the Se301 cells, which was verified by RTPCR. 5'/3' RACE analyses showed that the 3'- or 5'-end sequences of the viral transcripts are aligned to the host gene sequences in P8-Se301-C1 cells, suggesting that the SeMNPV genes may integrate into and be transcribed with the host genes in the P8-Se301-C1 cells. Furthermore, six additional viral gene transcripts, se11, se42, se44, se88, se91, and se127 (incorporated into chimeric fusion transcripts in the P8-Se301-C1 cells), were detected in the RACE analyses. Taken together, sixteen SeMNPV transcripts were identified in the P8-Se301-C1 cell strain. This study provides information to develop the understanding of baculovirus latent infections and superinfection exclusion.
P8 - Se301 - C1菌株的细胞是甜菜夜蛾细胞克隆,每个克隆都含有部分甜菜夜蛾多核多角体病毒(SeMNPV)基因组,并且对同源SeMNPV感染具有抗性。这些细胞不产生病毒后代,这表明该感染是一种潜伏样病毒感染。为了研究P8 - Se301 - C1细胞中所含的SeMNPV基因,对P8 - Se301 - C1细胞和甜菜夜蛾Se301细胞的从头转录组进行了分析和比较。从P8 - Se301 - C1细胞中总共获得了54,569,296条读数,产生了112,565个最终单基因,平均长度为1,093 nt。从Se301细胞中总共获得了56,865,504条读数,产生了102,996个最终单基因,平均长度为1,082 nt。通过RNA - Seq在P8 - Se301 - C1细胞中检测到10个SeMNPV基因转录本(se5、se7、se8、se12、se43、se45、se89、se90、se124和se126),而在Se301细胞中未检测到,这通过RT - PCR得到了验证。5'/3' RACE分析表明,在P8 - Se301 - C1细胞中,病毒转录本的3'端或5'端序列与宿主基因序列对齐,这表明SeMNPV基因可能在P8 - Se301 - C1细胞中整合到宿主基因中并与宿主基因一起转录。此外,在RACE分析中还检测到另外6个病毒基因转录本,se11、se42、se44、se88、se91和se127(在P8 - Se301 - C1细胞中整合到嵌合融合转录本中)。综上所述,在P8 - Se301 - C1细胞株中鉴定出16个SeMNPV转录本。本研究为深入了解杆状病毒潜伏感染和超感染排斥提供了信息。