Cloning Of Alfalfa Phytochrome A Gene Using Rapid Amplification Of CDNA Ends

In many countries,fall dormancy is the important index on spicies choice of alfafa, so the study of it is focused by scientists .it has been proved that the short day is main environmental factor causing alfafa fall dormancy ,that is showed that the alfalfa has photoperiodic effects and the phyA,phyB are the important photoreceptors.To study the gene of alfalfa is helpful to solve spicies fall dormancy choice in production and understand mechanism of fall dormancy .Because homology is very high between alfafa and legume, In the experiment , fabaceous phyA gene sequences were compared ,the result is that homology of these sequence exceed 90%,so a pair of primer was designed in homology region, a small segment sequence was amplified;it is proved that the small segment sequence belongs to alfafa phyA by NCBI BLAST,then depending on the known sequence, tow primers matched with outer and inner primer of 3′RACE and 5′RACE kit were designed , nested-PCR amplified production were cloned by TA clone .by sequencing and BLAST ,gene sequence has high homology with other legume phyA ,the result proves amplified gene are alfafa phyA. This cDNA fragment was 3130bp in length.