Double function epidermal growth factor and its preparation method and uses
An epidermal growth factor, dual-function technology, used in skin care preparations, chemical instruments and methods, skin diseases, etc., can solve the problem of non-specificity of damaged tissue, lack of specificity of wound tissue, and repair performance of gastrointestinal ulcers Poor and other problems, to achieve the effect of clear drug efficacy, no adverse side effects, and low cost
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Embodiment 1
[0024] Embodiment 1: Construction of PSL-CBD-EGF recombinant plasmid
[0025] 1. Using the primers P1 and P2 shown in Figure-1, the cDNA of L is used as a template to synthesize EGF PCR using high-fidelity TaqPCR enzyme.
[0026] serial number
name
Specification
unit
quantity
1
10× buffer
microliter
10
2
50mmol / L
microliter
4
3
P1 primer
5mm
microliter
2
4
P2 primer
5mm
microliter
2
5
cDNA
100ng
microliter
1
6
Taq enzyme
5u / ul
microliter
1
7
microliter
72
8
DNTP(10mm)
microliter
8
[0027] After preparing the above reaction system on an ice bath, centrifuge for 1 minute (3000 rpm) to mix the components in the system, and then place the sample in a PCR synthesizer. The conditions for the synthesis reaction a...
Embodiment 2
[0077] Example 2: Evaluation of CBD binding activity in EGF-CBD
[0078] In the 4 sets of experiments in Figure-3, the petri dishes were all coated with collagen, and then the test reagents were added:
[0079] A. Medium
[0080] B. Medium + CBD
[0081] C. Medium + EGF
[0082] D. Medium + EGF-CBD
[0083] Incubate at 37°C for 24 hours, then discard the culture medium in each plate, wash the culture plate with buffer solution (PBS) four times, and then detect with EGF antibody, the results show that: in group D, the CBD in EGF-CBD can Effectively bind to collagen molecules. At the same time, culture "NRK49F" cells with A.B.C.D four-group test-treated dishes. After inoculating the same number of cells, culture them at 37°C for 48 hours. Microscopic examination shows that EGF-CBD can Very effective in promoting cell growth.
Embodiment 3
[0084] Example 3: Comparison of EGF-CBD and EGF activity
[0085] Group 1: Medium
[0086] Group 2: Medium + CBD
[0087] Group 3: Medium + EGF
[0088] Group 4: culture medium + EGF-CBD
[0089] Figure-4 shows that each group was inoculated with the same number of cells (NRK49F). After culturing at 37°C for 48 hours, the growth rate of the cells is: EGF-CBD>EGF>CBD≥Medium, which shows that EGF-CBD can effectively promote cell growth like EGF.
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