Cloning And Expression Of The Pan-allergen Profilin Gene In Amaranthus Spinosus L. Pollen

Profilin has been described as a pan-allergen present in pollen of trees, grasses and weeds. Amaranthus spinosus is an Amaranthaceae, Amaranthus annual herb. Amaranthus spinosus. pollen is playing an important role in southern China which can cause severe anaphylaxis. So, it is of great theoretical and practical significance for us to clone and express the gene of this pan-allergen. In this paper, we firstly searched the known plant pollen profilin amino acid sequences from the Expasy database, then the bioinformatics method was used for sequence alignment and phylogenetic tree constructed, a pair of conservative primers was designed by homology comparison to the core sequence. At last, Two genes encoding Amaranthus spinosus pollen profilin were isolated by touchdown polymerase chain reaction(touchdown PCR) amplification, named prf7 and prf23. They were preliminary identify as a pan-allergen gene with sequencing and comparative analysis. Expression of the recombinant prf23 was carried out in E.coli using vector pET 44a, and it showed that prf23 could encode a protein according to SDS-PAGE identification. Finally, we analysed the three-dimensional structure and antigenicity between the two pan-allergens. The recombinant allergens obtained might be useful for the immunotherapeutic treatment and genetically engineered modification.