Coding

Part:BBa_K4279001

Designed by: Kong Yangyang   Group: iGEM22_SubCat_China   (2022-09-07)
Revision as of 08:21, 26 September 2022 by Lzn0220 (Talk | contribs)


W1-lipase

W1-lipase

Profile

Name: SP-lipase

Base Pairs: 1854 bp

Origin: Lactiplantibacillus Plantarum, genome

Properties: a lipase for triacylglyceride digestion.

Usage and Biology

BBa_K4279000 is the coding sequence of SP-lipase. Lipase is a primary lipase critical for triacylglyceride digestion in humans and is considered a promising target for the treatment of obesity [1]. Triacylglycerol lipase is the primary lipase secreted by the pancreas, and is responsible for breaking down dietary lipids into unesterified fatty acids (FAs) and monoglycerides (MGs). Medically, lipases are targets for therapeutic intervention in the treatment of obesity. The focus of applied research with lipases has been to exploit the unusual properties of lipolytic systems for the production of chiral pharmaceuticals, improved detergents, and designer fats [2]. Obesity is a medical condition in which excess body fat accumulates to the extent that it may have a negative effect on health, leading to reduced life expectancy and/or increased health problems. Diverse approaches to the prevention and treatment of obesity have been reported [3-5]. W1-lipase (EC 3.1.1.3) is a lipase amplified from Pseudomonas sp. 7323. The SP-lipase is made up of 265 aa [6].

Figure 1. The structural modeling of an extremophilic bacterial lipase isolated from saline habitats..

Construct design

1. Construction of the lipase expression plasmids The SP-lipase gene was amplified from the pseudomonas and then inserted in the XhoI and HindIII sites of pET28a (Figure 2).

Figure 2. W1-lipase and SP-ligase expression plasmids in this projec.

In order to build our plasmids, plasmid pET28a was digested with XhoI and HindIII (Figure 3), and we used T4 DNA ligase to ligate the fragments and the vector. Then we transformed the recombinant plasmids into E. coli DH5α competent cells and coated on the LB (Kanamycin) solid plates.

Figure 3. Gel electrophoresis of double-enzyme digested pET28a-SP-lipase.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 777
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 46
  • 1000
    COMPATIBLE WITH RFC[1000]


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