Construction of cDNA library of Dunaliella salina and cloning of kinesin like calmodulin-binding protein gene

Aim:To construct the cDNA library of Dunaliella salina and clone a new gene,kinesin like calmodulin-binding protein(KCBP).Methods:Total RNA was isolated and purified from Dunaliella salina. cDNA was then synthesized from isolated mRNA and ligated into the pAP3neo predigested vector. The recombinant plasmids were transformed into Escherichia coli DH10B by electroporation. Then the titers and recombinant rates were determined by the number of single clones.KCBP was screened from the cDNA plasmid library of Dunaliella salina by PCR.Results:The titer of the cDNA library was 5.6×106 pfu/mL, the recombination rate was about 90%, the sizes of most inserted fragments ranged from 0.4 to 6.0 kb with an average size of 1.9 kb. The screened gene KCBP belonges to kinesin-14 family.Conclusion:KCBP is cloned and screened successfully from the cDNA library of Dunaliella salina constructed in this study.