Method for observing distribution of protein homodimers on plant cell membrane
A homodimer, cell membrane technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, plant products, etc., can solve problems such as the difficulty of labeling homodimers, and achieve clear images, accurate results, Simple to use effects
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Embodiment 1
[0032] Example 1, Observation of the distribution of homodimers formed by the Arabidopsis HIR1 protein on the cell membrane
[0033] 1. Construction of HIR1 expression vector
[0034] The HIR1 gene was cloned and sequenced by PCR using high-fidelity Taq enzyme (Pfu). After the BiFC expression vector and the HIR1 gene fragment were digested and recovered with the same restriction endonuclease, the HIR1 gene (SEQ ID No.1) was respectively connected to the BiFC expression vector pNYE and pCYE with T4 DNA ligase, and the two recombinant expression vectors obtained were The vectors were named HIR1-nYFP and HIR1-cYFP, respectively.
[0035] The structure of the recombinant expression vector HIR1-nYFP is described as: the recombinant plasmid obtained after inserting the HIR1 gene (without stop codon) shown in SEQ ID No.1 between the restriction sites BamHI and KpnI of the pNYE vector.
[0036] The structure of the recombinant expression vector HIR1-cYFP is described as: the recombi...
Embodiment 2
[0046] Example 2, Observation of the distribution of homodimers formed by Arabidopsis Rem1.3 on the cell membrane
[0047] 1. Rem1.3 expression vector construction
[0048] The Rem1.3 gene was cloned and sequenced by PCR using high-fidelity Taq enzyme (Pfu). After the BiFC expression vector and the Rem1.3 gene fragment were digested and recovered with the same restriction endonuclease, the Rem1.3 gene (SEQ ID No.2) fragment was connected to the BiFC expression vector pNYE and pCYE respectively with T4 DNA ligase, The two obtained recombinant expression vectors were named Rem1.3-nYFP and Rem1.3-cYFP respectively.
[0049] The structure of the recombinant expression vector Rem1.3-nYFP is described as: the Rem1.3 gene (without stop codon) shown in SEQ ID No.2 is inserted between the restriction sites BamHI and KpnI of the pNYE vector to obtain recombinant plasmid.
[0050] The structure of the recombinant expression vector Rem1.3-cYFP is described as follows: the Rem1.3 gene (...
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