Plasmid used directly in intramuscular injection for treating diabetes
A technique of intramuscular injection and diabetes, applied in gene therapy, using vectors to introduce foreign genetic material, metabolic diseases, etc., can solve problems such as safety and problems
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[0030] 1 Construction of recombinant eukaryotic expression plasmid containing human CGRP gene
[0031] The eukaryotic expression plasmid pcDNA3.1(-)( figure 1 ) is commercially available from Invitrogen Corporation (Invitrogen Corporation, 1600 Faraday Ave. PO Box 6482, Carlsbad, CA92008, USA). The human CGRP gene is digested from the PTT42 plasmid, and the 844bp cDNA fragment containing the CGRP gene is recovered. It was connected to the downstream of the CMV promoter in the plasmid pcDNA3.1(-), and was controlled by the CMV promoter (see SEQ ID NO: 1 for the CGRP gene sequence). For the construction process of the recombinant plasmid, see figure 2 .
[0032] 2 Preparation of diabetic mouse model
[0033] Adopt classic method, select body weight 18-20 gram, the male C57BL mouse of 6-8 weeks, inject the streptozotocin (STZ) of 40mg / kg dose every day, inject continuously for 5 days, observe blood glucose and insulin concentration change ( Figure 3A and B). At 7 days, 2 ...
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