Humanized pika leptin protein, gene coding same and application thereof
A leptin protein, humanized technology, applied in the field of genetic engineering, can solve problems such as being in the blank
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Embodiment 1
[0048] Example 1 Artificially synthesized humanized pika leptin gene sequence
[0049] The sequence of SEQ ID No.1 was synthesized by artificial gene synthesis, and the gene synthesis was completed by Shanghai Sangon Bioengineering Co., Ltd.
Embodiment 2
[0050] Embodiment 2 Preparation of recombinant humanized pika leptin protein
[0051] 1. Experimental materials and solution preparation
[0052] (1) Expression vector and engineering bacteria
[0053] The expression vector used to express the recombinant pika leptin protein is pMAL-c2x (NewEngland Biolabs, NEB Company), and the engineering bacteria is Escherichia coli BL21 (DE3).
[0054] (2) Kits, tool enzymes and biochemical reagents
[0055] Restriction enzymes and Taq enzymes are products of Dalian Bao Bioengineering Co., Ltd., blood biochemical detection kits were purchased from Zhongsheng Beikong Biotechnology Co., Ltd., yeast extract (Yeast extract) and tryptone (Tryptone) were purchased from Oxford Co. Products, other antibiotics and biochemical reagents are products of sigma company.
[0056] (3) Protein purification chromatography column and its medium
[0057] All chromatographic media and their chromatographic columns are products of GE Healthcare, USA. The N...
Embodiment 3
[0090] Example 3 Application of humanized pika leptin protein in improving blood sugar and blood lipids
[0091] 1. Establishment of mouse model of hyperglycemia and hyperlipidemia
[0092] (1) Experimental animals
[0093] 40 male weaned (21 days) mice weighing about 25 g (purchased from the Experimental Animal Center of Hebei Medical University). Single-cage feeding, room temperature 18-25°C, relative humidity 50%-80%. 12 hours of light per day, free access to food and water. After 3 days of adaptation, blood was collected through the inner canthus, serum was separated, and baseline biochemical indicators were measured.
[0094] (2) High-fat feed formula: make containing lard 28%, white sugar 30%, milk powder 1%, egg yolk 18%, the high-fat feed of basic feed 33%.
[0095] (3) Establishment process of high-fat and high-glucose animal model
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