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以甘蓝型油菜隐性核不育两用系 S4 5 AB的可育株和不育株为材料 ,提取减数分裂期与单核期的花粉 m RNA,反转录合成 c DNA,并以其为模板扩增花粉特异基因 MS2 Bnap。结果减数分裂期能扩出产物 ,而单核期无任何产物。将减数分裂期的扩增产物克隆、测序。同源性分析表明 :在 S4 5 AB上克隆的 MS2 Bnap基因与 Gene Bank公布的拟南芥控制核育性基因 MS2和已报道的甘蓝型油菜的 MS2 Bnap基因的同源性分别为 88%和 99%。可育株 S4 5 B与不育株S4 5 A在 MS2 Bnap基因序列上有 4处碱基存在差异 ,其中 3处为同义突变 ,编码氨基酸未发生变化 :1处为错义突变 ,可育株为 AAA,不育株为 AGA。其三联体密码对应的氨基酸也发生了变化 ,AAA编码赖氨酸 ,AGA编码精氨酸。该变化可能与 S4 5 A的不育有关
Pollen m RNAs were extracted from the meiotic and monokaryotic pollen m RNAs of S4 5 AB inbred lines of recessive genic male sterile line of Brassica napus and reverse transcribed into c DNA. The pollen - specific gene MS2 Bnap was amplified by PCR. As a result, the product can be expanded during meiosis, without any product at mononuclear stage. The meiotic product was cloned and sequenced. Homology analysis showed that the homology between the MS2 Bnap gene cloned on S4 5 AB and the MS2 Bnap gene of Arabidopsis thaliana control nuclear fertility gene MS2 and reported Brassica napus was 88% and 99%. There were 4 bases in the sequence of MS2 Bnap between fertile plant S4 5 B and sterile plant S4 5 A, among which 3 were synonymous mutations, encoding amino acids did not change: 1 was missense mutation, fertile The strain is AAA, and the sterile strain is AGA. The amino acid corresponding to its triplet codon also changed, with AAA encoding lysine and AGA encoding arginine. This change may be related to S4 5 A infertility