荻MsFT1及MsFT2基因的克隆及序列分析
关键词 荻;光周期;Hd3a;FT
Key words Miscanthus Sacchariflor♀us;photoperiod;Hd3a;FT
1 材料与方法
1.1 试验材料及光周期ผ诱导
2 结果与分析
采用改良Trizol法提取荻叶片的总mRNA用琼脂糖凝胶电泳检测后,条带(图2)清晰完整性较好可以进行下一步。设计好的内参基因ACTIN11引物ACTIN11-F:CAAGCAGTAT GAAGATCAAGGT;ACTIN11-R:GCCTGACTCATCATACTCAC对反转后的cDNA进行PCR检测,均得到134 bp左右清晰的亮带,证明得到的cDNA第1条链结果良好,可进行下一步克 ッ隆的操作。cDNA作为扩增的模板,利用5对引物进行PCR扩增经凝胶电泳检测得到a、b、c、d、e共5条亮带,条带大小与预计大小相符。进行切胶回收纯化,载体连接,转入到DH5ฑa感受态扩增❣培养,再经过PCR检测为阳性菌后,每个PCR产物10个重复,送往海东盛生物有限公司进行测序。
3 讨论
4 参考文献
[5] HUANG T,BOHLENIUS H,ERIKSSON S,et al.The mRNA of the Arabidopsis gene FT moves from leaf to shoot apex and induces flowering[J].Science,2005,309(5741):1694-1696.
[6] WIGGE P A,KIM M C,JAEGER K E,et al.Integration of spatial and temporal information during floral induction in[J].Science,2005(309):1056-1059.
[7] CORBESIER L.FT protein movement contributes to long-distance signaling in floral induction of Arabidopsis[J].Science,2007(316):1030-1033.
[8] LIN M-K.Flowering locus T protein may act as the long-distance florigenic signal in the cucurbits[J].Plant cell,2007(19):1488-1506.
[16] BARDLEY D,CARPENTER R,COPSEY L,et al.Control of inflorescence architecture in Antirrhi-num[J].Nature,1996,379(6 568):791-797.