Difference between revisions of "Part:BBa K401000"

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Our PM2 was inserted into the pET28a vector and transformed into E. coli BL21 Star cells.  
 
Our PM2 was inserted into the pET28a vector and transformed into E. coli BL21 Star cells.  
 
We have to thank professor Yizhi Zheng and Yun Liu (College of Life Science, Shenzhen University. Guangdong, China) for sending us the plasmid with the gene, and, in consequence, let us develop our project.
 
We have to thank professor Yizhi Zheng and Yun Liu (College of Life Science, Shenzhen University. Guangdong, China) for sending us the plasmid with the gene, and, in consequence, let us develop our project.
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===Sequence===
 
===Sequence===

Revision as of 18:02, 27 October 2010

PM2 (Glycine max)


Function

PM2 corresponds to the LEA3 protein and it comes from soybean. LEA proteins are known for their roles in plant embryogenesis, as important dessication-resistance factors. It has also been shown that they confer tolerance under several stress conditions in Escherichia coli. You can read more about it in our wiki and in several references, as, for example, the following: Liu et al. (2010) Curr Microbiol 60: 373-378. PM2 can be useful as a general stress-resistance factor. It could improve survival under low or high temperatures, water stress and maybe other atypical conditions.


Original source

Our PM2 was inserted into the pET28a vector and transformed into E. coli BL21 Star cells. We have to thank professor Yizhi Zheng and Yun Liu (College of Life Science, Shenzhen University. Guangdong, China) for sending us the plasmid with the gene, and, in consequence, let us develop our project.


Sequence

The sequence of PM2 is the following one:


Then, in order to amplificate it and clone it into the pSB1C3, we used the following primer:

  • Forward actagtagcggccgctgcagATGGCGTCCAAGAAAC
  • Reverse tctagaagcggccgcgaattcTGCGTCTATATATAC

(capital letters indicate the region of the sequence that pairs with the coding sequence of PM2).


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 439
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 143
    Illegal BsaI.rc site found at 940
    Illegal BsaI.rc site found at 1006
    Illegal BsaI.rc site found at 1093
    Illegal SapI.rc site found at 1255