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The Research on Functional Genes of Cellulase by Biosensor

Author: YinJuan
Tutor: YuanXingZhong；ZengGuangMing
School: Hunan University
Course: Environmental Engineering
Keywords: Compost Biosensor Core-shell magnetic nanoparticles Reye Trichoderma Cellobiohydrolase enzyme
CLC: X705
Type: Master's thesis
Year: 2010
Downloads: 60
Quote: 0
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Abstract


Agricultural and industrial solid waste disposal is a very effective approach - micro-organisms in the compost, the process plays an important role. Tracking and monitoring of microbial populations composting system and the dynamic changes of the microbial functional gene analysis is conducive to in-depth understanding of the entire composting process microbiology mechanism of microbial inoculation and compost process innovation has a certain significance. Biosensing technology is a rapid, specific, real-time, online detection of emerging technology, it is the use of biometric role towards measuring material analysis determination of various biological groups, environmental pollutants and their degradation process involves inspecting object points or analysis of metabolite detection provides a new technology platform. This article focuses on tracking and monitoring of microbial populations function genes of composting system to carry out the gene-based sensing technology. DNA sensors developed to detect environmental efficient degradation of cellulose cellulase dynamic changes of functional genes, to overcome the traditional limitations of the analytical methods in environmental pollution control process real-time detection. Focused on a key enzyme in the degradation of cellulose - cellobiohydrolase enzymes (CBH). NCBI website reesei CBH Ⅱ gene sequence analysis of all sequence homology between the sequence using software based on sequence homology analysis to find functional genes exist segment, the application of probe design software functional genes The sequence segments of the design of hybridization probes and PCR primers. First multi-walled carbon nanotubes - the polymer conductive polymer composite films as well as Au-SiO2-Fe3O4 core-shell nanomaterials both nanomaterials as DNA vector. The carrier assembly with NH2 Reye Trichoderma cellobiohydrolase enzyme the type Ⅱ coding genes made of DNA sensors, and then with the Target chain, signal chain competitive hybridization. And carrier usage, ELISA dosage, hybridization time and hybridization conditions were optimized. DNA sensors based carboxylated multi-walled carbon nanotubes with polyethylene polymer substances (PEDOT) composite film. MWCNTs-PEDOT composite film modified electrode can be more effective to improve the conductivity of the electrode to enhance the sensitivity of the sensor, but multi-walled carbon nanotubes with carboxyl groups with NH2 reesei cellobiohydrolase lyase encoding type II gene crosslinking combination, can be combined with subsequent HRP enzyme labeled protein substances, the experiment exists interference still need further improvement. Nano-particle size is moderate dispersion control, biocompatibility, high, and the synthetic low cost advantages, the use of gold nanoparticles in situ amplification technology made each a gold / silicon / iron three composite core-shell magnetic nanoparticles developed into a competition-style hybrid electrochemical nano identify CBHII gene DNA sensor, and the lower limit of detection of 10-13M; and depth strain genome extraction, PCR technology to extract the cellulase CBHII coding genes, and the extracted DNA strands used in the core-shell type Further discussion of of magnetic nanomaterials DNA carrier DNA sensor.

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CLC: > Environmental science, safety science > Processing and comprehensive utilization of waste > General issues > Solid waste disposal and utilization
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