Building body of mediating plant fertility and application thereof
A construct and plant technology, applied in the field of constructs mediating plant fertility, can solve the problems of low sorting efficiency and poor effect
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no. 1 example
[0112] The first embodiment, transformation of maize plants with recombinant expression vector DBN100123 comprising male fertility gene Ms45, DAM methylase gene and herbicide resistance gene PAT
[0113] 1. Construction of recombinant cloning vectors DBN01-T, DBN02-T, DBN03-T, DBN04-T
[0114] The synthesized 5126:Ms45 fragment was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01-T, which was constructed Process such as figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; 5126 is the corn anther specific promoter (SEQ ID NO:1); Ms45 is the coding sequence of maize male fertility gene (SEQ ID NO:2); MCS is t...
no. 2 example
[0166] Second embodiment, transformation of maize plants with recombinant expression vector DBN100010 comprising male fertility gene Ms26, α-amylase gene and herbicide resistance gene PAT
[0167] 1. Construction of recombinant cloning vectors DBN05-T, DBN06-T, DBN07-T, DBN08-T
[0168] The synthesized 5126:Ms26 fragment was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN05-T, which was constructed Process such as image 3 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; 5126 is the corn anther specific promoter (SEQ ID NO:1); Ms26 is the coding sequence of maize male fertility gene (SEQ ID NO:16); MCS is the mul...
no. 3 example
[0220] The third embodiment, transformation of rice plants with recombinant expression vector DBN100011 comprising male fertility gene DPW, α-amylase gene and herbicide resistance gene PAT
[0221] 1. Construction of recombinant cloning vectors DBN09-T, DBN10-T, DBN11-T, DBN12-T
[0222] The synthesized gDPW fragment was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN09-T. The construction process is as follows: Figure 5 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of bacteriophage f1; LacZ is the LacZ start codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; gDPW is the rice male fertility Genome sequence (SEQ ID NO:23); MCS is multiple cloning site).
[0223] Then the recombinant cloning vector DB...
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