Indicator gene used in TRV-mediated gene silencing system as well as construction method and application of carrier thereof

A technology of gene silencing and gene carrier, applied in the field of molecular biology, to achieve the effects of long-lasting traits, simple construction, and significant silencing phenotypes

Inactive Publication Date: 2015-04-01
ZHENGZHOU UNIV
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  • Abstract
  • Description
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ZDS Gene as an indicator gene of TRV-mediate...

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  • Indicator gene used in TRV-mediated gene silencing system as well as construction method and application of carrier thereof
  • Indicator gene used in TRV-mediated gene silencing system as well as construction method and application of carrier thereof
  • Indicator gene used in TRV-mediated gene silencing system as well as construction method and application of carrier thereof

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Experimental program
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Embodiment Construction

[0023] 1. ZDS Gene homologous cloning and construction of ZDS-TRV VIGS vector

[0024] Search by NCBI ZDS Gene sequence, design conservative primers at ATG and TGA. The common tobacco safflower Dajinyuan cDNA was used as a template for PCR amplification, the target fragment was recovered, T-A ligated, transformed into competent Escherichia coli (DH5α), cultured overnight at 37 ℃, and positive clones were obtained through colony PCR identification. Positive plasmid extraction, final sequencing to obtain common tobacco ZDS The full-length sequence of the coding region. get ZDS The full-length sequence of the coding region (CDS) is 1785 bp, encoding a total of 594 amino acids. ZDS The base sequence of the coding region is shown in SEQ ID NO:1. will sequence the correct ZDS Gene sequences and other plants ZDS Gene sequences are compared to find the conserved region, which is selected in the present invention ZDS The 633 bp gene fragment of 782-1414 at the 5′ end ...

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Abstract

The invention discloses an indicator gene used in a TRV-mediated gene silencing system, and the base sequence of the indicator gene is shown in SEQ ID NO:1. Virus induced gene silencing in the invention is a good tool for studying plant gene functions by using reverse genetics; according to the invention, an extremely good indicator gene carrier is provided for a tobacco rattle virus induced gene silencing technology; and the indicator gene carrier is simple in construction, significant in silencing phenotype and lasting in trait expression, and achieves a good indication effect on quick gene function researches.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to an indicator gene carrier, construction method and application. Background technique [0002] Virus-induced gene silencing (VIGS) is an important method for studying plant gene functions. It is simple, fast and efficient, does not require complex genetic transformation of plants, and can conduct functional research on lethal genes. . In recent years, a variety of VIGS vectors have been developed and widely used in various plants. Among them, tobacco rattle virus (TRV) is currently the most widely used VIGS vector. It is composed of two RNA virus strands, TRV1 and TRV2. TRV1 is used to assist TRV2 carrying target gene fragments to move in plants. As a viral vector, TRV has many advantages, such as mild virus symptoms, high and long-lasting silencing efficiency, and various tissues can produce silencing, so it is widely used. In particular, the improved tobacco rattle virus vect...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N15/83C12N15/66A01H5/00
Inventor 杨冉史艳梅王燃屈凌波林福呈罗朝鹏李锋魏春阳杨军
Owner ZHENGZHOU UNIV
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