Part:BBa_K3991000

ArsD

Profile

Properties: Gene technology for protecting patented bacterial strains

ArsD is a trans-acting repressor of the arsRDABC operon that confers resistance to arsenicals and antimonials in Escherichia coli.

Experimental approach

Production, purification, and sequcing analysis of recombinant ArsD-amilGFP

Figure 1: Gel Electrophoresis Results of PCR of amilGFP Genes..

Figure 1 shows gel electrophoresis results of amilGFP PCR. Column M is a 2K marker ladder. Columns 1-6 are PCR products of amilGFP genes All 1-6 columns displayed successful results at 700bp which could be used for later experiments. 

Figure 2: Gel Electrophoresis Results of PCR of ArsD Gene..

Figures 2 show the result for colony PCR identification on the E.coli with ARSD/amilGFP inserted that were cultivated previously. The purpose is to examine whether the E.coli contains expected gene segment of ARSD.

Function Tests

Function Test With NaH2PO4, Attempt 1

Such function tests were performed under the hypothesis that NaH2PO4 is structurally similar to arsenic compounds that were meant to be detected. NaH2PO4 is used to substitute for arsenic compounds due to experimental safety. No fluorescence was detected in all samples, indicating that the concentrations of NaH2PO4 may be too low or that genetically engineered E. coli will not react to NaH2PO4.

Function Test With NaH2PO4, Attempt 2

In order to verify our assumption in attempt 1, NaH2PO4 was used again in this testing, with greater concentration than the last test. No fluorescence was detected in all samples, indicating the inability of the genetically engineered E. coli to react with NaH2PO4.

Function Test With C2H6AsNaO2

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]