applied micro-fluidic biochips in systems and synthetic biology

1
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576 S541 scale-up production of (-CGTase and other enzymes by recombi- nant E. coli. doi:10.1016/j.jbiotec.2010.09.889 [PUB-O.29] Applied micro-fluidic biochips in systems and synthetic biology Jie Zeng Benjoe Institute of Systems Bio-Engineering, China Keywords: STFD; Micro-fluidic Chips; Systems Biology; Synthetic Biology Abstract: The design of bio-molecular systems within cell, it is a recently achievement of synthetic biology. Due to the structurity theory (Zeng, 1996) of synthetic biosystems and applied sys- tem genetics Bangzhe, 1994: 1).structure integration, cordinative organization of genes network in genome; 2).function adaptation, duplication (amplify) and specific expression of genome involved in cell signalling; 3).construct stratification, multiple stratum of genome constructed by transpose, recombination and amplifi- cation. We developed the techniques of the Sequence Tagged Fragments Display (STFD) for analysis of genes differential expres- sion between cell types, which will be very useful in systems medicine (Zeng, 1995) and systems bioengineering (Zeng, 1994). The genetic modified cell or bionic designed cell is an bio-molecular nano-machine, which can be used as cell nano-factory, or used as cell nano-computer. A kind of cell machine of bio-molecular com- plexity, it is constructed of molecules module, ADP-ATP energy system and information system of genes. To design signal trans- duction networks and synthesis gene regulatory structures of cells, we applied the technological integration of micro-fluidic biochips worked for drug discovery, isolation of non-known sequence gene, and design of cellular bioreactor or micro-nano systems of cells communication. References Zeng B.J. Structure theory of self-organization systems, Communications on Trans- genic Animals, CAS, No. 8-10, 1996. Bangzhe Jie Zeng, Transgenic animal expression system – the goldegg plan, Commu- nication on Transgenic Animals, CAS, Nov. 1994.(On the mendelian, molecular and system genetics, and the term “system genetics” was coined). Zeng B.J. From positive to synthetic medical science, Communications on Transgenic Animals, CAS, Nov. 1995. Zeng B.J. On the concept of systems biological engineering, Communications on Transgenic Animals, CAS, June, 1994. doi:10.1016/j.jbiotec.2010.09.890 [PUB-O.30] Evaluation of Two Real Time RT-PCR Methods for Norovirus GI and GII Detection in Live Bivalve Molluscs E. Suffredini , L. Cozzi, G. Ciccaglioni, L. Croci Istituto Superiore di Sanità - dipertimento di Sanità Pubblica Veteri- naria e Sicurezza Alimentare, Italy Keywords: Norovirus; shellfish; PCR Shellfish are recognized as a potential vehicle of viral diseases, however, because of the lack of reference methods for viruses detection, the European Regulations still relies on bacteriological parameters to establish their microbiological quality. The aim of the present study was the evaluation of the abil- ity of two Real Time RT-PCR methods (an in-house method and a commercial kit) on detecting Norovirus belonging to GI and GII, in shellfish. The analyses have been performed both on a Norovirus Reference Panel (NRP), constituted by various synthetic RNA of NoV GI, GII and GIV, and on mussels (Mytilus galloprovincialis) from class A and B harvesting areas. In the experiments carried out on NRP the in-house Real Time RT-PCR method provided better results. The analysis of the results (McNemar test) showed a statistically significant difference between the two methods, because of the inability of the commer- cial kit to detect most of the GI NoV. Among 90 shellfish analysed 34.4% were positive for NoV and 9.0% of these samples contained both NoV GI and GII. The two methods provided only a few dis- cordant results and because of the contemporary presence of both genogroups (GI and GII) in the same samples it was not possible to highlight significant differences. Both methods resulted suitable for the detection of NoV contamination in shellfish, however the in- house Real Time RT-PCR method presents the advantage to directly differentiate GI and GII, that should be important to deepen the knowledge on the circulation and the resistance on the environ- ment of the two genogroups. doi:10.1016/j.jbiotec.2010.09.891 [PUB-O.31] Bioengineering of methanotrophic consortia for reduction of methane emission in coal mines Xin-Hui Xing 1,, Hao Jiang 1 , Peixia Jiang 1 , Chong Zhang 1 , Yin Chen 2 , Colin J. Murrell 2 1 Tsinghua University, China 2 the University of Warwick, United Kingdom Keywords: methane; Coal Mines; Methanotrophs; Microbial Con- sortium Methanotrophs have promising applications in removal of CH 4 from the coal mines for the control of green-house gas and the safety management. Methanotrophic community in a coal mine and their enrichment process for biofilter construction as well as on site applications for methane reduction in coal beds are of great importance. In this research, based on the previous analysis of the methanotroph community in an alkaline coal mine in China (Han et al., 2009), CH 4 and artificial mash gas were used to domesticate the mixed methanotrophic cultures capable of stably and effectively abating CH 4 , respectively. Denaturing gradient gel electrophoresis (DGGE) was adopted to analyze the changes of microbial commu- nity structure during the enrichment process. Biofilters and the on site removal of methane from coal bed by using the enriched methanotrophic consortia were studied. Clone library and DGGE showed significant changes in micro- bial communities during the enrichment processes with different gases. By using the stably enriched methanotrophic consortium, the high cell density culture was achieved and the removal efficiency of CH 4 by the biofilter and spraying the consortium in the simu- lated coal bed reached more than 90%. This study demonstrated that the methanotrophic consortium enriched from the coal mine soils could be effective for reduction of methane control in coal mines. This work was supported by 863 Project of MOST of China (No. 2006AA02Z203) and the National Key Basic Research Program (973 plan) (No. 2007CB109203).

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Page 1: Applied micro-fluidic biochips in systems and synthetic biology

Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576 S541

scale-up production of (-CGTase and other enzymes by recombi-nant E. coli.

doi:10.1016/j.jbiotec.2010.09.889

[PUB-O.29]

Applied micro-fluidic biochips in systems and synthetic biology

Jie Zeng

Benjoe Institute of Systems Bio-Engineering, ChinaKeywords: STFD; Micro-fluidic Chips; Systems Biology; SyntheticBiology

Abstract: The design of bio-molecular systems within cell, it is arecently achievement of synthetic biology. Due to the structuritytheory (Zeng, 1996) of synthetic biosystems and applied sys-tem genetics Bangzhe, 1994: 1).structure integration, cordinativeorganization of genes network in genome; 2).function adaptation,duplication (amplify) and specific expression of genome involvedin cell signalling; 3).construct stratification, multiple stratum ofgenome constructed by transpose, recombination and amplifi-cation. We developed the techniques of the Sequence TaggedFragments Display (STFD) for analysis of genes differential expres-sion between cell types, which will be very useful in systemsmedicine (Zeng, 1995) and systems bioengineering (Zeng, 1994).The genetic modified cell or bionic designed cell is an bio-molecularnano-machine, which can be used as cell nano-factory, or used ascell nano-computer. A kind of cell machine of bio-molecular com-plexity, it is constructed of molecules module, ADP-ATP energysystem and information system of genes. To design signal trans-duction networks and synthesis gene regulatory structures of cells,we applied the technological integration of micro-fluidic biochipsworked for drug discovery, isolation of non-known sequence gene,and design of cellular bioreactor or micro-nano systems of cellscommunication.

References

Zeng B.J. Structure theory of self-organization systems, Communications on Trans-genic Animals, CAS, No. 8-10, 1996.

Bangzhe Jie Zeng, Transgenic animal expression system – the goldegg plan, Commu-nication on Transgenic Animals, CAS, Nov. 1994.(On the mendelian, molecularand system genetics, and the term “system genetics” was coined).

Zeng B.J. From positive to synthetic medical science, Communications on TransgenicAnimals, CAS, Nov. 1995.

Zeng B.J. On the concept of systems biological engineering, Communications onTransgenic Animals, CAS, June, 1994.

doi:10.1016/j.jbiotec.2010.09.890

[PUB-O.30]

Evaluation of Two Real Time RT-PCR Methods for Norovirus GIand GII Detection in Live Bivalve Molluscs

E. Suffredini ∗, L. Cozzi, G. Ciccaglioni, L. Croci

Istituto Superiore di Sanità - dipertimento di Sanità Pubblica Veteri-naria e Sicurezza Alimentare, ItalyKeywords: Norovirus; shellfish; PCR

Shellfish are recognized as a potential vehicle of viral diseases,however, because of the lack of reference methods for virusesdetection, the European Regulations still relies on bacteriologicalparameters to establish their microbiological quality.

The aim of the present study was the evaluation of the abil-ity of two Real Time RT-PCR methods (an in-house method and a

commercial kit) on detecting Norovirus belonging to GI and GII,in shellfish. The analyses have been performed both on a NorovirusReference Panel (NRP), constituted by various synthetic RNA of NoVGI, GII and GIV, and on mussels (Mytilus galloprovincialis) from classA and B harvesting areas.

In the experiments carried out on NRP the in-house RealTime RT-PCR method provided better results. The analysis of theresults (McNemar test) showed a statistically significant differencebetween the two methods, because of the inability of the commer-cial kit to detect most of the GI NoV. Among 90 shellfish analysed34.4% were positive for NoV and 9.0% of these samples containedboth NoV GI and GII. The two methods provided only a few dis-cordant results and because of the contemporary presence of bothgenogroups (GI and GII) in the same samples it was not possibleto highlight significant differences. Both methods resulted suitablefor the detection of NoV contamination in shellfish, however the in-house Real Time RT-PCR method presents the advantage to directlydifferentiate GI and GII, that should be important to deepen theknowledge on the circulation and the resistance on the environ-ment of the two genogroups.

doi:10.1016/j.jbiotec.2010.09.891

[PUB-O.31]

Bioengineering of methanotrophic consortia for reduction ofmethane emission in coal mines

Xin-Hui Xing 1,∗, Hao Jiang 1, Peixia Jiang 1, Chong Zhang 1, YinChen 2, Colin J. Murrell 2

1 Tsinghua University, China2 the University of Warwick, United KingdomKeywords: methane; Coal Mines; Methanotrophs; Microbial Con-sortium

Methanotrophs have promising applications in removal of CH4from the coal mines for the control of green-house gas and thesafety management. Methanotrophic community in a coal mineand their enrichment process for biofilter construction as well ason site applications for methane reduction in coal beds are of greatimportance. In this research, based on the previous analysis of themethanotroph community in an alkaline coal mine in China (Han etal., 2009), CH4 and artificial mash gas were used to domesticate themixed methanotrophic cultures capable of stably and effectivelyabating CH4, respectively. Denaturing gradient gel electrophoresis(DGGE) was adopted to analyze the changes of microbial commu-nity structure during the enrichment process. Biofilters and theon site removal of methane from coal bed by using the enrichedmethanotrophic consortia were studied.

Clone library and DGGE showed significant changes in micro-bial communities during the enrichment processes with differentgases. By using the stably enriched methanotrophic consortium, thehigh cell density culture was achieved and the removal efficiencyof CH4 by the biofilter and spraying the consortium in the simu-lated coal bed reached more than 90%. This study demonstratedthat the methanotrophic consortium enriched from the coal minesoils could be effective for reduction of methane control in coalmines.

This work was supported by 863 Project of MOST of China (No.2006AA02Z203) and the National Key Basic Research Program (973plan) (No. 2007CB109203).