Blood sugar reducing method of orally taking human insulin-like growth factor-I produced in silkworm bioreactor
A silkworm baculovirus and bioreactor technology, applied in the fields of genetic engineering and biopharmaceuticals, can solve the problems of low expression, poor activity, and poor post-processing of products, and achieve high expression rate and good biological activity.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0049] Example 1. Construction of recombinant virus rBmNPV-(IGF-I) and its expression in silkworm.
[0050] 1. Construction of recombinant transfer vector pVL1393-(IGF-I)
[0051] The IGF-I gene was digested with BamHI to pGEM3Z (IGF-I) plasmid to obtain the IGF-I / BamHI fragment, which was ligated with the pVL1393 vector digested with BamHI, transformed into TGI strain, and recombinants were screened therefrom. Further use the specific primers on the polyhedrin gene promoter sequence on the designed pVL1393 vector and the specific primer sequences on the IGF-I gene to carry out PCR reactions to verify the insertion direction of IGF-I, the construction of the vector and PCR identification like figure 1 and 2 shown.
[0052] 2. Construction of recombinant baculovirus rBmNPV-(IGF-I)
[0053] Inoculate approximately 1×10 6 Cells at 15cm 2 In the culture flask, after the cells adhere to the wall, remove the medium containing fetal bovine serum (FBS), wash with medium without ...
Embodiment 2
[0076] Embodiment 2, the oral hypoglycemic effect of IGF-I produced by the silkworm bioreactor.
[0077] The experimental animals used in this experiment were Kunming mice.
[0078] 1. Preparation of hyperglycemic mice: Feed the mice with 15% sucrose water instead of distilled water, and follow the method of "Medical Experimental Zoology" (Edited by Wei Hong, Sichuan Science and Technology Press, 1998) for other feeding methods.
[0079]2. After feeding with high-sugar water for 10 days, use the phosphomolybdic acid colorimetric method (edited by the Biochemical Teaching and Research Office of Peking University, biochemical experiment guidance. Higher Education Press, 1979) to measure the blood sugar concentration, and monitor the blood sugar content in the mouse blood after feeding sugar. The change.
[0080] 3. After feeding high sugar water continuously for 15 days, the blood sugar of the mice reached a certain concentration. Orally fed IGF-I hypoglycemic experiment. The...
Embodiment 3 I
[0083] Embodiment three, the relation of IGF-1 oral dosage and hypoglycemic
[0084] The preparation of all experimental animals, blood sugar measurement methods, and hyperglycemic mice is the same as in Example 2.
[0085] The experiment was divided into 6 groups, with 5 male and 5 mice in each group. Experimental group 1 is the normal mouse control; Experimental group 2 is high-sugar mice, which are fed silkworm blood with corresponding protein content by perfusion method as high-glucose control; Experimental group 3 is the high-sugar mice fed 2 μg IGF-I group by perfusion method; Experimental group 4 is the high-sugar rats fed with 4 μg IGF-I group; Experimental group 5 is the high-sugar rats fed with 6 μg IGF-I group; Experimental group 6 is the high-sugar rats fed with 8 μg IGF-I group . Sixteen hours after IGF-I treatment, blood samples were taken to measure the blood glucose concentration in the mice of each group. Experimental results such as Figure 7 Shown:
[0...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More