Amplification, Sequencing and Phylogenetic Analysis of EF-1 Gene in Apis Family (Hymnoptera: Apidae)

摘要

In this study, with the help of Elongation Factor (EF-1) sequences reported in Apis family analyzed the relationship of the same family in Tamilnadu, South India, India, through nucleotide amplification, sequencing and Phylogenetic tree analysis. The worker honey bee specimens were collected from the colonies in different locations of Tamilnadu, South India. Total Genomic DNA was extracted from worker bees thorax region by using slight modification of high salt extraction protocol and the extracted Genomic DNA were subjected to PCR amplification done by using nuclear protein gene EF-1 primers. Amplified products were analyzed in 1% analytical agarose gel electrophoresis and the molecular weight of the amplified product size of amplicon is approximately ~1100 bp. The amplified product of Apis cerana indica (MSS2) were sequenced and submitted in Genbank (GU935342). Phylogenetic analysis done with other four species sequence data of EF-1 gene collected from Genbank. The phylogenetic tree was constructed by using tree top software program from GeneBee service was started with a set of aligned sequence clustal W. The tree shows clearly that the strain MSS2 is highly cluster with Apis florea and Apis cerana with 100% bootstrap value. Also strain MSS2 had maximum sequence identity (99%) with Apis niqrocincta and Apis cerana than Apis dorsata (96%) and Apis florea (95%). It is assuring that strain MSS2 had a maximum identity 99% and phylogenetically cluster with Apis cerana indica alone. EF-1 analysis of honey bee is an effective tool to classify the honey bee species.