A myb gene regulating cotton fiber elongation and its application
A cotton fiber and gene technology, applied in the field of MYB family transcription factors and their coding genes, can solve the problems of lagging research on regulatory mechanisms and functions that have not been systematically reported
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[0020] 1. Isolation and cloning of cotton GhFEM1 genome DNA sequence and cDNA
[0021] Cotton fiber RNA was extracted 9 days after flowering by CTAB method (performed by Li XB, Cai L, Cheng NH, Liu JW, 2002.Molecular characterization of the cotton GhTUB1 gene that preferentially expressed in fiber.Plant Physiol.130:666-674), with SuperScript TM II RNase H-Reverse Transcriptase (Invitrogen Life Technologies) was reverse transcribed into cDNA; then, using cDNA as template, gene-specific primers (GhFEM1-ORF-L 5'-GGGGAATTCATGGATGTAACAAGCACACC-3' and GhFEM1-ORF-R 5' -CTTGGATCCTCACTCGATCTTACTCATCC-3') and high-fidelity DNA polymerase PFU, 94°C-5 min, 1 cycle; 94°C-1 min, 58°C-1 min, 72°C-1.5 min, 29 cycles; finally, 72 Under the PCR reaction conditions of elongation at ℃ for 10 minutes, a DNA fragment of about 1 kb (FEM1 gene) was obtained by amplification, as shown in SEQ ID No. 1. This fragment was inserted into the pBluescript SK-vector. Use the T3 and T7 promoters on the pla...
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