基于甘蔗花叶病毒的基因沉默在……中
Sugarcane Mosaic Virus-Based Gene Silencing in .
作者信息
Ali Sajed, Ahmad Nasir Idrees, Rafiq Muhammad, Javed Butt Shahid, Ihsan Farooq, Qayyum Rao Abdul, Husnain Tayyab
机构信息
Department of Biotechnology, School of Sciences, University of Management and Technology, Sialkot Campus, Sialkot, Pakistan.
Centre of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
出版信息
Iran J Biotechnol. 2017 Dec 29;15(4):260-267. doi: 10.15171/ijb.1536. eCollection 2017.
Potyvirus-based virus-induced gene silencing (VIGS) is used for knocking down the expression of a target gene in numerous plant species. virus (SCMV) is a monopartite, positive single strand RNA virus. pBINTRA6 vector was modifi ed by inserting a gene segment of SCMV in place of (TRV) genome part 1 (TRV1 or RNA1) and the two nonstructural proteins of TRV2(RNA2). : SCMV construct was inoculated into 3-4 weeks plant leaves either by using a needleless syringe or applying pricking with a toothpick. The construct (SCMV-RNA2) successfully induced post-transcriptional gene silencing (PTGS) of the target genes and I through agroinoculation proving that SCMV is a substitute of the RNA1, which plays a pivotal role in the systemic gene silencing. 2-3-weeks of post inoculation, target genes' silencing was observed in the newly developed noninoculated leaves. The newly developed construct expresses the knocked down of the endogenous as well as exogenous genes and only four weeks are required for the transient expression of the gene silencing based on SCMV-VIGS system.
基于马铃薯Y病毒属病毒的病毒诱导基因沉默（VIGS）被用于在多种植物物种中敲低靶基因的表达。甘蔗花叶病毒（SCMV）是一种单分体、正义单链RNA病毒。通过插入SCMV的一个基因片段取代烟草脆裂病毒（TRV）基因组第1部分（TRV1或RNA1）以及TRV2（RNA2）的两个非结构蛋白对pBINTRA6载体进行了改造。具体做法是：使用无针注射器或用牙签穿刺的方式将SCMV构建体接种到3至4周龄的植物叶片中。该构建体（SCMV-RNA2）通过农杆菌接种成功诱导了靶基因的转录后基因沉默（PTGS），证明SCMV可替代在系统基因沉默中起关键作用的RNA1。接种后2至3周，在新长出的未接种叶片中观察到了靶基因的沉默。新构建体可使内源性和外源性基因表达被敲低，基于SCMV-VIGS系统的基因沉默瞬时表达仅需四周时间。
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