Difference between revisions of "Part:BBa K1509002"

Latest revision as of 01:22, 18 October 2014

Mainly responsible for the flocculation of Bacillus sp. F2

It is responsible for the flocculent ability of Bacillus sp. F2 and has been converted to E.coli DH5α. It has been proved that strain of E.coli positive clone FC2 could express flocculent activity, with the flocculent efficiency of 90%, thus can be a good choice in environmental applications. The 1071bp gene consists of 29%T, 18%C, 25%G and 29%A.

Usage and Biology

The Bacillus sp. F2 that was used in this study was stored at -40°C in 20% glycerol. The bacteria from the stock cultures were pre-cultured in Luria-Bertani culture medium (LB) prior to use.DH5α was taken as the host for recombinant plasmids. The pET-28b (+) was prepared as an overexpression vector to produce the target protein. Rosetta pLysS was used as the host for expression of the flocculation gene under the control of the T7 promoter. E. coli transformants were grown at 37°C in LB medium.

Result

For comparative purposes, two E.coli strains with or without flocculation gene were used. The results were shown in Fig. 1. It was showed that the control strain could not settle efficiently within a short time. After 20 min static placement, only 55% of cells were flocculated. In contrast, the recombinant strain was strongly flocculent, 70% of cells were flocculated within only 10 min. At last, about 80% of the cells were settled after 20 min. The results indicated that the flocculation gene was expressed successfully in the E.coli strain. The flocculation ability of the recombinant strain could give us a new sight to remove cells from liquid environment. However, we only observed the flocculation characters of the strain in phosphate buffer containing 4 mM Ca²⁺, and we did not test the flocculation ability of the strain in the waste water which contains various metal ions and organic pollutant, and that is what we will focus on in our further study.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1029
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1029
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1029
Illegal BamHI site found at 1008
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1029
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1029
1000
COMPATIBLE WITH RFC[1000]

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 ===Usage and Biology===
-The ''Bacillus sp.'' F2 [1] that was  used in this study was stored at -40°C in 20% glycerol. The bacteria from  the stock cultures were pre-cultured in Luria-Bertani culture medium (LB) prior to  use.''DH5α'' was taken as the host for  recombinant plasmids. The pET-28b (+) was prepared as an overexpression vector  to produce the target protein. ''Rosetta  pLysS'' was used as the host for expression of the flocculation gene under  the control of the T7 promoter.'' E. coli'' transformants were grown at 37°C in LB medium.
+The ''Bacillus sp.'' F2 that was  used in this study was stored at -40°C in 20% glycerol. The bacteria from  the stock cultures were pre-cultured in Luria-Bertani culture medium (LB) prior to  use.''DH5α'' was taken as the host for  recombinant plasmids. The pET-28b (+) was prepared as an overexpression vector  to produce the target protein. ''Rosetta  pLysS'' was used as the host for expression of the flocculation gene under  the control of the T7 promoter.'' E. coli'' transformants were grown at 37°C in LB medium.
 [[File:ReNEFU_CHINA_fa_fig1.png]]
 ===Result===
-For comparative purposes, two E.coli strains with or without flocculation gene  were used. The results were shown in Fig. 1. It was showed that the control  strain could not settle efficiently within a short time. After 20 min static placement, only 55% of cells were flocculated. In contrast, the  recombinant strain was strongly flocculent, 70% of cells were flocculated  within only 10 min. At last, about 80% of the cells were settled after 20 min.  The results indicated that the flocculation gene was expressed successfully in  the E.coli strain. The flocculation  ability of the recombinant strain  could give us a new sight to remove cells from liquid environment. However, we  only observed the flocculation characters of the strain in phosphate  buffer containing 4 mM Ca<sup>2+</sup>, and we did not test the flocculation ability of the strain in the waste water which contains  various metal ions and organic pollutant, and that is what we will  focus on in our further study.
+For comparative purposes, two ''E.coli'' strains with or without flocculation gene  were used. The results were shown in Fig. 1. It was showed that the control  strain could not settle efficiently within a short time. After 20 min static placement, only 55% of cells were flocculated. In contrast, the  recombinant strain was strongly flocculent, 70% of cells were flocculated  within only 10 min. At last, about 80% of the cells were settled after 20 min.  The results indicated that the flocculation gene was expressed successfully in  the ''E.coli'' strain. The flocculation  ability of the recombinant strain  could give us a new sight to remove cells from liquid environment. However, we  only observed the flocculation characters of the strain in phosphate  buffer containing 4 mM Ca<sup>2+</sup>, and we did not test the flocculation ability of the strain in the waste water which contains  various metal ions and organic pollutant, and that is what we will  focus on in our further study.
 [[File:ReNEFU_CHINA_fa_fig2.png]]
-===References===
-.   Fang Ma, Junliang Liu, Shugeng  Li, Jixian Yang, Liqiu Zhang, Bo Wu, Yanbin Zhu.''Development of complex microbial flocculant''.  China water and wastewater, 2003, 19(4):1-5.