USTC/SuXiaofeng

From 2007.igem.org

Xiaofeng Su(Allen Su)-Undergraduate of Cellular and Molecular Biology in School of Life Sciences, USTC, P.R. China

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.

My project description by visual comics: All we need are the specific artificial 'repressor fish' that can definitely bite the specific 'operator hook' exclusively

My Work Basis-By Redesigning lac-repressor and operator. This figure comes from 'Roberto Kopke Salinas, etc. ChemBioChem 2005, 6, 1628 – 1637'

• 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

1. 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.

1. 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.

• Key Results