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Zinc lipoprotein of giardia-resistant action target and medical application

A zinc lipoprotein and giardia technology, applied in the field of anti-parasites, can solve the problems that there are no ideal drugs and vaccines

Inactive Publication Date: 2021-03-16
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the control of giardiasis still mainly depends on the use of anti-giardiasis drugs, such as metronidazole, etc., but due to the emergence of drug resistance, there is no ideal drug and vaccine for controlling giardiasis so far

Method used

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  • Zinc lipoprotein of giardia-resistant action target and medical application
  • Zinc lipoprotein of giardia-resistant action target and medical application
  • Zinc lipoprotein of giardia-resistant action target and medical application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The giardia zinc lipoprotein (zinc-finger domain (ZFD) containing protein, ZFD) disclosed by the present invention has the molecular formula: C 923 h 1438 N 274 o 295 S 15 , with a relative molecular mass of 21.6KD and an isoelectric point of 6.70, which can regulate the activity of Giardia telomerase and thus play an anti-Giardia effect. SWISS-MODEL is used to predict the 3D structure of ZFD. Its gene sequence is as follows: Shown in SEQ No 1, the result is as figure 1 shown.

Embodiment 2

[0021] TRAP detects the effect of ZFD ribozyme on telomerase activity:

[0022] After 10 μL of ZFD ribozyme was electrotransfected into Giardia trophozoites, the telomerase activity of Giardia ribozyme transfection group, negative control group and empty vector group was detected by TRAP method. After the trophozoites were washed 3 times with RNase-free PBS, 1×10 7 Add 200 μL of lysate to each worm, and grind 35 times on ice with a high-speed tissue grinder, 1-2 sec each time. Lyse in an ice bath for 20 minutes, then centrifuge at 12000g at 4°C for 15 minutes, take the supernatant, measure the protein concentration by BCA method, aliquot, react at 30°C for 30 minutes, and inactivate at 95°C for 5 minutes. Then 1 μL of PR primers and 1 μL of rTaq were added for PCR amplification. The amplification parameters were: 94°C for 1 min, 59°C for 1 min, 72°C for 1 min, and 33 cycles. The reaction results were detected by electrophoresis. The results showed that there was no signifi...

Embodiment 3

[0024] RT-PCR method to detect the effect of ZFD ribozyme on telomere length:

[0025] After extracting Giardia RNA and reverse-transcribing it into cDNA, telomere primers and internal reference were designed according to the repeat sequence of Giardia telomeric DNA, and PCR amplification was carried out at 94°C for 5 min; 94°C for 30 s, 60°C for 30 s, 72°C for 1min, repeat 30 cycles; 72°C for 10min; take it out after cooling down to 4°C, and verify the PCR results by electrophoresis.

[0026] In order to detect whether the result of the telomere primer meets the standard, after the RT-PCR is finished, 5 μL of the PCR product is taken for agarose gel electrophoresis detection. The results showed that after 30 cycles of amplification, multiple bands appeared from about 60bp, and the length extended to more than 500bp. A reliable method for telomere length, electrophoretic verification of PCR results (as attached image 3 shown).

[0027] The telomere length of Giardia experi...

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PUM

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Abstract

The invention provides zinc lipoprotein of a giardia-resistant action target and medical application. Protein ZFD capable of remarkably inhibiting giardia propagation is found, and the protein ZFD acts on giardia TRBD to inhibit giardia propagation, reduce the telomerase activity and influence the telomere length; and a new target ZFD is provided for treatment of giardia, and a foundation is laidfor drug intervention of giardia disease in the future. Finding of the action target for telomerase by new telomerase related protein is a new way for development of anti-giardia drugs.

Description

technical field [0001] The invention provides a zinc lipoprotein of an anti-giardia action target, as a new anti-giardia action target, which can be used to develop anti-giardia drugs, and belongs to the technical field of anti-parasites. Background technique [0002] Giardia duodenum ( Giardia duodenalis ), referred to as Giardia (Giardia), as a zoonotic parasite, mainly parasites in the small intestine of humans and some mammals, causing giardiasis with diarrhea as the main feature, and seriously endangering the health of humans and other animals . At present, the control of giardiasis still mainly depends on the use of anti-giardiasis drugs, such as metronidazole, etc., but due to the emergence of drug resistance, there is no ideal drug and vaccine for controlling giardiasis so far. Because Giardia can be "immortalized" through repeated passages in vitro, its regulation is closely related to telomeres. Telomere is a special nucleoprotein complex located at the end of e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/44A61K38/17A61P33/02
CPCA61K38/00A61P33/02C07K14/44
Inventor 张西臣郑敬彤李建华宫鹏涛张楠李新王晓岑杨举马赫然
Owner JILIN UNIV
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