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	'''(C)'''I've develop the selection work by the means of Blue White Selection on top agar Luria-Bertani broth to obtain the R-P pairs.Below are some pictures from my experiment design and experiment work.		'''(C)'''I've develop the selection work by the means of Blue White Selection on top agar Luria-Bertani broth to obtain the R-P pairs.Below are some pictures from my experiment design and experiment work.
-	[[Image:USTC_allen10.jpg]] [[Image:USTC_allen7.jpg]]	+	[[Image:USTC_allen10.jpg|thumb|left|300px|Figure.7.My arranged selection work desciption]] [[Image:USTC_allen7.jpg|thumb|right|200px|Figure.8.Selection Results-the Red-Mark the colony are a target]]
-	[[Image:USTC_allen8.jpg]] [[Image:USTC_allen9.jpg]]	+	[[Image:USTC_allen8.jpg|thumb|center|300px|Figure.9.retest of my results of P-R pairs]]
	*;Key Results		*;Key Results

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Revision as of 17:14, 20 October 2007

XIAOFENG SU

Undergruduate Student of Cellular and Molecular Biology,USTC

Email: allensue@mail.ustc.edu.cn (preference) or xiaofsu@gmail.com

Phone: +86-551-3602469 (Lab)

Mobile: +86-13866722084

Address: Room 439, School of Life Sciences, USTC, Hefei, Anhui, P.R.China, 230026

Research Interest

• Directed Evolution for Seeking New Protein-DNA Interactions

• Approaches of Synthetic Biology for Forming Novel "Genetic Engineering Machines"

• Differentiation and Development of Stem Cells

Research Work

• Overall Description

For obtaining the signaling transduction parts of repression with high fidelity, I've experimentally designed and acquired some specific repressor-promoter pairs(R-P pairs or P-R pairs)based on Lactose Operon by directed evolution on plate. Besides, through quantitative assay,the novel artificial R-P pairs I selected have been tested for their binding performance so that P-R pairs of highest affinity and specificity can make a figure out of P-R pair candidates. Most of the parts in my work have been BioBricks-Standardized and work as BioBrick parts.

• Experimental Design

1. Construction of Expression Library of Lac-Repressor Family [Collaboration]
2. Synthesis of Promoter Sequence with Specific Operators
3. Construction of Low-copy Reporter System with Specific Opertors
4. Selection of Promoter-Repressor Pair(P-R Pair) including re-testing validity of these combination
5. Transfering the Operators to Double Reporter Systems [Collaboration]
6. Quantitative assay of Repression Intensity and Specificity of P-R Pairs
7. Results From RM(Repression Matrix) to ORM(Orthogonal RM)

• Work Process and Summary

(A)The Directed Evolution must be conducted on the base of 2 components-the repressor system and promoter-reporter system.By collaborations with other team members, I've successfully establish these two systems shown as below.

(B)Within the work of establishment of two selection systems, a time consuming step is synthesis of target specific promoters that contain the mutated site design by Do. On account of economic and convenient aspect, I use 2-Step unparallel PCR by 3 fragments of primers and 2 times of PCR with different reaction conditions.As Below.

(C)I've develop the selection work by the means of Blue White Selection on top agar Luria-Bertani broth to obtain the R-P pairs.Below are some pictures from my experiment design and experiment work.

• Key Results