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Formulation Development for Microbial Flocculant HBF-3 and Genetic Transformation of vip3Aa into Paecilomyces Lilacinus

Author: ZhaoNi
Tutor: YuZiNiu
School: Huazhong Agricultural University
Course: Microbiology
Keywords: Microbial flocculant Freeze-drying Concentrated by rotary evaporation Vegetative insecticidal protein from Paecilomyces lilacinus
CLC: Q93
Type: Master's thesis
Year: 2011
Downloads: 26
Quote: 0
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Abstract


This study were two independent HBF-3 : a microbial flocculant dosage forms flocculant HBF-3 is a deep sea salt Aeromonas V3a '(Halomonas sp. V3a') an acidic polysaccharide, has good flocculation activity . This experiment for the further development and utilization of bio- flocculant dosage form study . As a protective agent, 5 % sucrose fermentation broth made ??of solid dosage forms by freeze-drying technology lyophilized HBF-3 capable of quick recovery water-soluble flocculating activity of 82.4% , maintaining a high flocculating activity . Concentrated by using a rotary evaporator , the concentrated solution was diluted after flocculation activity compared to the initial fermentation broth , little change. Add 1.0g / L potassium sorbate , liquid concentrated flocculant samples at 4 ° C , 28 ° C , 37 ° C , 50 ° C storage for one month , be able to keep more than 90% of the flocculation activity . 2 vip3Aa gene on lilac Paecilomyces genetic transformation lilac Paecilomyces (Paecilomyces lilacinus) as the root-knot nematode ( Meloidogyne spp . ) Cyst nematode (Heterodera spp.) Plant pathogenic nematode hyperparasite , is considered is the most promising plant pathogenic nematode biocontrol fungi one . Vip3Aa soluble protein expression in the vegetative stage of Bacillus thuringiensis with high virulence of the beet armyworm and black cutworm . This is the first the vip3Aa genes into lilacinus genome to both prevention and control root-knot nematode while combating engineered strains of black cutworm and other soil pests . The the vip3Aa cloned into pET28b () vector and expressed in E. coli after get Vip3Aa proteins purified polyclonal antibody for this antigen to immunize New Zealand rabbits were preparing anti- Vip3Aa . PcDNA3.1 (-) , and pcDNA3.1 ( ) respectively intermediate plasmid construct the genetic vip3Aa transformed with the plasmid of Agrobacterium tumefaciens the transformation vector puPVNT and LB4404 . G418 resistance gene as a selectable marker , ATMT (Agrobacterium tumefaciens-mediated transformation) method vip3Aa gene lilac to Paecilomyces 9410 strains genetic transformation . PCR confirmed the vip3Aa gene was successfully integrated into the genome of the 9410 strain Paecilomyces lilacinus .

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CLC: > Biological Sciences > Microbiology
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