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Study on Bacterial Ghosts of Haemophilus Parasuis Based PhiX174 Gene E Lysis Cassette-mediated

Author: WangMin
Tutor: ZhuBiFeng;YangXuFu
School: Nanchang University
Course: Biochemistry and Molecular Biology
Keywords: Parasuis Bacterial ghost Vector construction
CLC: S852.5
Type: Master's thesis
Year: 2007
Downloads: 27
Quote: 0
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Abstract


Bacterial ghost (BG) is a novel adjuvant nature vaccine or DNA vaccine delivery system . It is the removal of the cytoplasm of non-active , but contains the cell morphology and antigenic structure of the characteristics of the bacterial cell envelope . It is achieved by cleavage of the expression of genes in Gram - negative bacteria , genetic inactivation . Bacteria specter performance of the natural function of the cell surface and the antigenic structure . Therefore , the technology used in animal vaccine research , new safe and effective vaccine has great significance for the development of infection prevention for animal diseases . Pig leather Rather 's disease pathogens parasuis recipient strain recombinant plasmid containing PhiX174 E lysis gene , located thermal λpL/pRcI857 lacpo - lacI8 promoter repressor system and tol expression by cloned into the plasmid vector of the system control . E -mediated lysis thermostat mainly through the λpR gene expression regulatory pathway for the thermal stability of the extension of the the λpR promoter or cI857 repressor system . ΛpR promoter or operon region mutation lead to a new expression system, i.e. the repressor E gene in the expression of 37 ℃ induction of cell lysis within the range of 39 to 42 ° C , but allow . Main results are as follows ; 1 will be transformed into E. coli DH5α containing plasmid pHH43 lysis gene E , ??the induced expression and found that the cracking rate of the lysis genes in E. coli can be achieved up to 99.84% , observed by a scanning electron microscope ( SEM ) in bacteria of the poles , and the bacteria split position to form a hydrophobic pore . But the pHH43 transformed parasuis did not succeed. 2 was cloned into the expression cassette (Cassette) PHIX174 gene E pLS88 shuttle plasmid constructed the pLS88-E plasmid . By restriction enzyme digestion and sequencing theory PHIX174 E gene . The recombinant plasmid pLS88-E were transformed into E. coli and Haemophilus parasuis induced expression comparison cracking rate . The results showed that the two bacteria induced temperature-sensitive point showed a decline in cell concentration decrease in plate culture living cells ; parasuis cracking rate of 99.84% , the rate is about 82.04% of E. coli lysis . Electron microscopy revealed cell holes in the bacterial membrane ; This proved parasuis ghost successful preparation . These results laid the technical foundation for the development of effective parasuis the specter vaccine and other gram-negative bacteria and bacterial ghost vaccine .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Serological
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