【摘要】 根据Genbank登陆的紫花苜蓿,抗寒基因cas18(L07516)的mRNA序列设计引物,提取低温胁迫下紫花苜蓿叶片中的RNA,以反转录的cDNA为模板,进行PCR反应,扩增产物经回收、连接、转化和酶切鉴定为阳性重组子,其序列长度为755bp。经比较,与Genbank登陆的紫花苜蓿cas18基因序列的同源性达到80%以上,表明所克隆的基因序列为紫花苜蓿cas18基因部分序列。低温胁迫下,紫花苜蓿中cas18基因的表达量在供试的7个品种间存在显著的差异,不同品种cas18基因编码区也存在部分序列上的差异,推测这可能是导致不同品种抗寒能力差异的原因。更多还原

【Abstract】 Total RNA in leaves of alfalfa cultivars under low temperature stress was extracted for resersing transcription into cDNA and then the cDNA was as a template for PCR reaction according to designing primer of cds of Medicago sativa cas18 cold-regulated gene (L07516). The amplified products were positive recon identified by using procedures of recovery, connection, transformation and enzyme separation. The length of cloned gene sequence was 755bp Identity reached 80% with Medicago sativa cas18 gene, which means that the cloned gene sequence was the partial cas18 gene of alfalfa. The expression amount of gene cas18 is different with 7 alfalfa cultivars under cold stress. AH,LM and ZD are higher, ZM is middle, the expression of other 3 cultivars is similar, but lower. There are some differences in cas18 cds among 7 cultivars that may lead to the cold stress different in 7cultivars.更多还原