The coding region of CnFatB1, CnFatB2, and CnFatB3 of coconut was cloned in E.coli DH5? by heat shock method using pMD18-T. After identifying validated positive clones, FatB gene family was sequenced. The nucleotide sequence of CnFatB1 was related to C.nucifera acyl-ACP thioesterase FatB1 with a nodal value of 99, CnFatB2 showed that was related to C. nucifera acyl-ACP thioesterase FatB2 with a nodal value of 97 and CnFat3 was related to C. nucifera acyl-ACP thioesterase FatB3 with 98 for a nodal value respectively. Expressed vector was constructed by ligating CnFatB1, CnFatB2 and CnFatB3 to linearized pCAMBIA 1302 and transformed into E.coli by heat shock. Moreover, expression vector was also transformed into Agrobacterium tumefaciens GV3103. Potential colonies harboring FatB genes were screened by colony PCR, and confirmed by using expression vector as template for PCR and by enzyme digestion using BglII and BstEII. All results showed that CnFatB1, CnFatB2 and CnFatB3 were in expected size.