Chinese cabbage fertility relevant gene and method of using the gene in constituting artificial male sterility line of Chinese cabbage and similar crops
A male sterile line and genetic technology, applied in genetic engineering, botany equipment and methods, biochemical equipment and methods, etc., can solve crop self-incompatibility instability, difficulty in parental reproduction, time-consuming Power and other issues
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[0016] Take the construction of 'Shanghai Qing' common cabbage and 'Youqing' cabbage as an example.
[0017] Obtaining Antisense Fragments of Fertility Related Genes
[0018] According to the full-length sequence (SEQ ID No.1) of the Chinese cabbage fertility-related gene, primers were designed between the 901st base and the 1338th base in the gene, and the 438bp sequence was amplified as an antisense gene fragment (SEQ ID No.2). And according to the multiple cloning restriction site of plant binary vector pBI121 ( figure 1 ), respectively design two PCR amplification primers containing BamH I and Xba I restriction endonuclease sites and three protective bases, the sequence of which is: P1 5'-GCT GGA TCC ACC ATT AGG TTT TTT AGA CA- 3'; P2 5'-GATTCT AGA GCC ACT AAC AAA CTT CCC TG-3'. The PCR amplification conditions were: 94°C, pre-denaturation for 3min; 94°C, denaturation for 1min; 50°C, annealing for 45s; 72°C, extension for 45s; 72°C, extension for 10min; storage at 4°C; ...
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