Cloning And Analysis Of Skip Homologous Gene CDNA From Medicago Falcatal

In this research, we cloned and analysised the cDNA sequence of SKIP homologous gene in Medic ago faleata L. Under 1/2 MS media supplemented with 20% PEG 6000, the Medicago falcata L. seedings involved in drought stress. As splicing factor, the MfSKIP cDNA was isolated by RT-PCR and homology-based cloned from Medicago falcata L. seedings. Analysis show that full-lenghth of the cDNA is 2256 bp, it has a open reading frame of 1836 bp, which coded 611 amino acid. And the putative protein with a predicted molecular weight of 69.1 kDa and PI 8.78. The sequence homology analysis indicated MfSKIP cDNA sequence exhibited 96% identity respectively with that from Medicago truncatula. The deduced amino acid sequence from the nucleotide sequence of the cDNA showed considerable homology with known members of the SNW_SKIP protein family. The S-N-W-K-N motif, which is absolutely conserved in SNW_SKIP protein family, was also found in MfSKIP protein. At the same time, MfSKIP protein amino acid sequence was analysised on the secondary structure, signal peptide, hydrophilicity and hydrophobicity, Phosphorylation sites, transmembrane domain. It turned out that this protein is hydrophilic protein and locted in the nucleus and have no transmembrane structure and signal peptide. In addition, this protein secondary structure is mainly composed of random coil. Analyzing the phylogenetic tree, we found MfSKIP has a closely genetic relationship with Medicago truncatula and Cicer arietinum. Furthermore, we cloned and analysised the MfSKIP from genome. Sequence analysis showed that the gene have no introns.