Identification and application of cotton fibre-specific HB (homeobox) transcription factor gene KNAT7
A gene and gene coding technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of the size of the HB gene and the difference in the structure of the encoded protein.
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[0040] 1. Isolation and cloning of full-length cDNA of cotton KNAT7 gene
[0041] A new gene of the KONX family was isolated from the cotton fiber cDNA library (according to Li XB et al., 2002, Molecular characterization of the cotton GhTUB1 gene that is preferentially expressed in fiber.Plant Physiol 130: 666-674), named as KNAT7.
[0042] 2. Cloning of the full-length DNA sequence of the KNAT7 gene
[0043] Design a pair of primers (see Table 1) at both ends of the KNAT7 gene ORF, use cotton genomic DNA as a template, at 94°C-5 minutes, 1 cycle; 94°C-1 minute, 58°C-1 minute, 72°C-2 Minutes, a DNA fragment of about 1 kb was obtained under 32 cycles of PCR reaction conditions, and the fragment was inserted into the PBS-T vector. The target DNA fragments were sequenced using the T3 and T7 promoters on the PBS plasmid. The obtained sequence was analyzed by DNAstar software, compared with the cDNA sequence, and the size and insertion position of the intron were analyzed.
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