An Investigation Of Spiders Endosymbiont Of Chengdu Plain And Preliminary Screening Of Penicillin Acylase From It

Spiders with their endosymbionts are a pool that consists of enormous secondary metabolites and enzymes. At the gate of the third wave of biocatalysts, many kinds of new thoughts, new methods new technologies concerning about biocatalysts development emerged. Three main representative novel resources realm including the ocean, endosymbiontic system, and extreme environmental microbes attracted the crowds’ attention. But spider endosymbionts researches and exploitationes just fall behind. This work was carried forward under such circumstance.Ordinary specieses of spider captured outdoor were used for endosymbionts diversity survey and microbes profile emerged, which implied universal discrepancy. Next generation sequencing experiment was done for the purpose of deepening research results. The thesis produced 2063 OTUs which affiliated to 27 phylums, 60 classes, 77 orders, 167 families and 453 genera. Another work based on 16 S library was done for the puopose of exploring the existence of target microbe genus. As far as we known, this paper was a relatively comprehensive work in spider endosymbiont taxonomy.A common spider species( Nephila clavata) was used as start material, spider endosymbiont metagenome was extracted and corresponding library was constructed(total library capacity reached 15 k meanwhile coverage surpassed 10 folds). DOP-PCR combined with Touch Down method were applied to screen penicillin acylase core sequence.A sequence shared partial homology with the penicillin acylase(EC.3.5.1.11) harbored by Pseudomonas mosselii SJ10 was retrieved. The identity between it and the known ones was 85% and 88% regarding to nucleotide and amino acids respectively. Sequence analysis implied it was a novel penicillin acylase.Based on core sequence, thermal asymmetric interlaced PCR experiment was designed to screen the whole arrayed library in this paper expecting for obtaining complete sequences of the target. These efforts failed in this paper, which call on further deeper researches.