High-efficiency transgenic cotton expression vector and application thereof

A technology for plant expression vectors and genes, applied in plant expression vectors and its application fields, can solve problems such as not being suitable for cotton transgenics, low accuracy in screening cotton transgenic positive plants, difficulty in obtaining research results, etc., to achieve easy and accurate screening Effect

Inactive Publication Date: 2011-02-02
CHINA AGRI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, after years of research, it has been found that spraying Basta is not very accurate for screening cotton transgenic positive plants. At the same time, because many domestic research departments adopt the pollen tube channel transgenic me

Method used

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  • High-efficiency transgenic cotton expression vector and application thereof
  • High-efficiency transgenic cotton expression vector and application thereof
  • High-efficiency transgenic cotton expression vector and application thereof

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Embodiment 1

[0036] Example 1, construction of convenient screening and high-efficiency transgenic cotton expression vector

[0037] 1. Acquisition of high-efficiency transgenic cotton expression vector

[0038] The pSPT vector is modified on the basis of pCambia1300. The promoter of the target gene we use is a Super strong promoter, and the Flag sequence is added behind the promoter. In addition, the reporter gene is replaced by the tfdA gene from the Hygromicin(R) gene. The final vector was named pSPT ( figure 2). The construction process of the pSPT vector is as follows:

[0039] The CaMV35S promoter in pCAMBIA1300 was replaced with the Super strong promoter. Firstly, our laboratory modified the multiple cloning site of pCAMBIA1300. The modified sites are: SalI, KpnI, BamI, SpeI, SalI, ApaI, SmaI, SwaI, PstI, HindIII, XbaI and AccIII. We first synthesized DNA fragments containing MfeI, BamI, SpeI, SalI, ApaI, SmaI, SwaI, PstI, HindIII, XbaI, AccIII and HindIII restriction sites, th...

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Abstract

The invention discloses a plant expression vector and application thereof. The plant expression vector comprises an expression box expressing a tfdA gene, wherein the expression box comprises a CaMV35S promoter, a tfdA gene and a terminator that are sequentially connected; the nucleotide sequence of the tdfA gene is a sequence 2 in a sequence table, and the nucleotide sequence of the CaMV35S promoter is a sequence 1 in the sequence table. The construction of the vector of the invention has very important application values on the construction of the gene recombinant expression vector of the cotton, the gene transformation (especially providing incontestable evidence for the transgenes of pollen tubes in China), later generation screening of the transgenes and the whole cotton gene engineering for the research of the cotton transgenes.

Description

technical field [0001] The invention relates to a plant expression vector and its application, in particular to a high-efficiency transgenic cotton expression vector and its application. Background technique [0002] The earliest typical example of transgenic animals is the "super mouse" reported by Palmiter et al. of Washington University in 1982. But before that, in 1974, Jaenisch et al introduced the DNA of SV40 into the embryo sac of mice by microinjection, and detected the DNA of SV40 in the liver and kidney tissues of the mice, which proved that the external It is possible to introduce the source gene into embryonic cells and achieve integration. In 1980, Gordon and others successfully introduced the DNA fragments of herpes virus and SV40 into the mouse genome for the first time by microinjection of fertilized egg pronuclei. At that time, Gordon and Ruddle called the transformed mice "transgenic mice". [0003] Transgenic plants. In 1983, Agrobacterium-mediated metho...

Claims

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Application Information

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IPC IPC(8): A01H5/00C12N15/82
Inventor 齐俊生巩志忠陈智忠王妍卿罗祖勇张定朋
Owner CHINA AGRI UNIV
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