Tmem52 protein for regulating glycolipid metabolism, coding gene, sgRNA and application of Tmem52 protein
A technology for glucose and lipid metabolism and protein, applied in the field of medicine, can solve the problems of prohibitive surgery risks, and achieve the effect of regulating abnormal glucose and lipid metabolism and reducing body weight
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[0033]In the present invention, the preparation method of the Tmem52 protein preferably includes: protein cloning and / or tissue extraction; the tissue preferably includes blood and / or pancreas. The method for cloning the protein is not particularly limited in the present invention, and methods well known to those skilled in the art can be used.
[0034] In the present invention, the obesity-related diseases preferably include: one or more of metabolic syndrome, diabetes and fatty liver. The Tmem52 protein provided by the present invention can significantly reduce the body weight of mice, regulate and regulate abnormal glucose and lipid metabolism through the Tmem52 protein provided by the present invention, and can be used for the treatment of obesity and related diseases.
Embodiment 1
[0037] Cloning and purification of the Tmem52 protein consisted of the following steps:
[0038] 1) Acquisition of the target DNA fragment (design primers and amplify the DNA fragment with the sequence shown in SEQ ID NO.2);
[0039] 2) Construct recombinant plasmid;
[0040] Step 1) and step 2) are completed by Gemma Gene;
[0041] 3) introducing the recombinant plasmid into Escherichia coli for expression, purification and identification; the specific steps are as follows:
[0042] Take 100 μL of competent Escherichia coli BL21(DE3), add 4 μL of the recombinant plasmid constructed in step 2) and bathe in ice for 30 minutes; heat shock at 42 °C for 45 s, and bathe in ice for 1 min; add 700 μL of LB medium, shake at 37 °C for 45 min; plate and grow overnight;
[0043] Pick a single colony in 50ml LB liquid medium, shake the bacteria overnight; take 10ml of the bacterial solution and inoculate it into a large bottle containing 1L of LB medium, shake until the OD value is 0.6-...
Embodiment 2
[0048] A sgRNA targeting gene Tmem52 (shown in SEQ ID No.2), the sgRNA is composed of sgRNA-L and sgRNA-R; the nucleotide sequence of the sgRNA-L is shown in SEQ ID No.3, The nucleotide sequence of the sgRNA-R is shown in SEQID No.4.
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