Utilization Of PiggyBac Transposon For Honeybee Transgenic Research

Honeybee was a new model organism,with the completion and further improvement of honeybee genome sequencing,how to clarify the function of these genes was significant to exploit and apply the significant theoretical gene and economic effective gene.this paper refered to utilization of piggyBac transposon for honeybee transgenic research.In this study,we analyzed potential endogenous transposons of Apis cerana cerana and Apis mellifera using bioinformatics methods.It turned out to be one transposon element protein in Apis mellifera(Gene ID: 100576623)and one transposon element protein in Apis cerana cerana(GenBank: PBC25285.1),the former was about 1799 bp,coding 294 aa,the latter was about 579 bp,coding 192 aa.Transcriptional analysis was initially performed to verify its transposition activity.In this study,three kinds of transposition plasmids from professor Martin Beye in the laboratory of Evolutionary Genetics at the university of Duesseldorf,Germany,were used to enzyme digestion and sequencing to verify basic structure information.In addition,exogenous plasmids were injected into Western honeybee eggs to evaluate the transcription activity of their plasmids.The target band was detected by reverse transcription PCR and it was confirmed that the plasmid was indeed expressed in honeybee eggs.Based on the above research,This paper discussed the gene transfer function of the piggyBac vector in the Apis mellifera.PCR amplification and sequencing analysis showed that the piggyBac transposon can integrate the exogenous Rubia red fluorescent protein gene into the honeybee genome.And weak red fluorescent protein expression was detected in the microinjection group.This study confirmed that the piggyBac transposon system can implant exogenous genes into the honeybee genome.