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
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[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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