[Cloning and bioinformatic analysis of IspE and IspH genes in Lonicera japonica and its substitutes].

OBJECTIVE: To clone IspE and IspH unigene from Lonicera japonica and its substitutes, and analyze their gene sequence, protein properties and transcriptional level.

METHOD: IspE and IspH unigene ware obtained from the transcriptome dataset of L. japonica. Full-length cDNA of IspE and IspH were cloned from buds of L. japonica, L. japonica var. chinensis, L. hypoglauca and L. dasystyla using RT-PCR technology, and named as LJIspE, LHIspE, LJCIspE, and LDIspE, LJIspH, LJCIspH, LHIspH and LDI-spH, respectively. And we also predicted the structure and function of IspE and IspH proteins.

RESULT: IspE contained an open reading frame that consisted of 1 221 bp, encoding one polypeptide with 422 amino acids. A complete open reading frame of IspH gene consisted of 1 380 bp and encoded 459 amino acids. Both IspE and IspH ware non-secreted proteins and localized in the chloroplast. Transcripted level of IspE and IspH in bud of L. japonica, L. hypoglauca and L. dasystyla was not significantly difference, but their transcripted level in L. japonica var. chinensis was significantly higher than that in L. japonica.

CONCLUSION: The clone of IspE and IspH will help for further research on the synthesis of terpenes, aroma and color.