Difference between revisions of "Part:BBa J100034"

(→‎Promoters and Their Primers)
(→‎Promoters and Their Primers)
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groe-down  CGGGGTACCTGAAATAACCTCCTCAATAGTATGA            KpnI
 
groe-down  CGGGGTACCTGAAATAACCTCCTCAATAGTATGA            KpnI
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[HERE SHOULD BE A FIGURE]
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<P>Figure 1. pMAA0911H-ASN transformation vector</P><br>
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<P>The promoters Groe (BBa J100034), SecA (BBa K1469002), and P43 (BBa K208002) were used to replace the original promoter hpall in WB0911H-ASN respectively. Here ASN stands for L-asparaginase (EC.3.5.1.1). By the comparison of the ASN activity expressed in the transformants, we found that the p43 promoter was the strongest promoter. Therefore, it was selected for our project.
  
 
=== Strains and Plasmids ===
 
=== Strains and Plasmids ===

Revision as of 15:58, 14 October 2019

groE promoter

The default state of this promoter is off and can be induced by heat. It is 44 base pairs long and is located 72 bases upstream of the start site of groE. The groE gene is from E. coli.

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]

Promoters and Their Primers

Several promoters with high expression level that are commonly used in Bacillus subtilis were screened to determine the most efficient ones for our experiments on the construction of biological parts. Three of such promoters, Groe (BBa J100034), SecA (BBa K1469002), P43 (BBa K208002), exist in the iGEM registry already.Here we only show the data of Groe(BBa_J100034).

The primer sequences for individual promoters used for our synthetic biology project are as follows:

groe-up CGCGGATCCCCAATACTGTTTTCTCAAATGGTATGTA BamHI

groe-down CGGGGTACCTGAAATAACCTCCTCAATAGTATGA KpnI

[HERE SHOULD BE A FIGURE]

Figure 1. pMAA0911H-ASN transformation vector


The promoters Groe (BBa J100034), SecA (BBa K1469002), and P43 (BBa K208002) were used to replace the original promoter hpall in WB0911H-ASN respectively. Here ASN stands for L-asparaginase (EC.3.5.1.1). By the comparison of the ASN activity expressed in the transformants, we found that the p43 promoter was the strongest promoter. Therefore, it was selected for our project.

Strains and Plasmids

The bacterium strain we used was B. subtilis WB600, and the plasmid was WB0911H-ASN (with the antibiotic ampicillin and kanamycin resistance cassette). Both the strain and plasmid were kindly donated to us by Professor Jian CHEN at Jiannan University, China.