Cloning and identification of the full-length cDNA sequence of FLA8 gene from Dunaliella salina

Aim:To get the full-length cDNA sequence of FLA8 gene from Dunaliella salina(D.salina).Methods:A pair of degenerated primers was designed according to conserved homologous amino acid sequences of GYNGTIF and NEDPKDA of kinesin-2 motor subunit of FLA8 from microalgae and other organisms.The full-length cDNA of FLA8 gene was obtained by RT-PCR,3'and 5'RACE technology.Results:The results showed that the full-length cDNA sequence of the cloned FLA8 gene was 2 612 bp,possessing a 90 bp 5'UTR,a 167 bp 3'UTR and a 2 355 bp open reading frame encoding 784 amino acids.The theoretical pI value of FLA8 was 6.65 and the relative molecular weight was 85 097.Amino acid sequence homologous analysis indicated that FLA8 of D.salina shared high homology with other organisms,such as Volvox carteri f.nagariensis(98%),Chlamydomonas reinhardtii(99%),Micromonas(92%).Conclusion:The full-length cDNA sequence of FLA8 gene from D.salina has been cloned.