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Crystal of thymidylate synthetase (ThyX) of helicobacter pylori and preparation method

A technology of thymidylate synthase and Helicobacter pylori, applied in the fields of molecular biology and applied microbiology, can solve the problems of undiscovered public reports

Inactive Publication Date: 2010-11-24
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] Through document search, do not find the public report of the same document as the content of the present invention

Method used

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  • Crystal of thymidylate synthetase (ThyX) of helicobacter pylori and preparation method
  • Crystal of thymidylate synthetase (ThyX) of helicobacter pylori and preparation method
  • Crystal of thymidylate synthetase (ThyX) of helicobacter pylori and preparation method

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Experimental program
Comparison scheme
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Embodiment Construction

[0034] Reagent

[0035] Primers Shanghai Sangong

[0036] Endonuclease Takara

[0037] Hampton Research Kits Hampton USA

[0038] instrument

[0039] AKTA purifier Hiload 16 / 60 Superdex200 prep grade GE

[0040] Ultrafiltration Concentrator Millipore, USA

[0041] Centrifuge Hitachi, himac CRT, rotor R5S2

[0042] Ni-NTA column 5ml Novagen

[0043] The ThyX gene was cloned from the gene of Helicobacter pylori 26695, cloned into the expression vector pET-30a, and after amplified culture in LB liquid medium, a large amount of thymidine was secreted and expressed in the medium after 4 hours of induction acid synthase. Then, the cells were crushed and purified by affinity chromatography and molecular sieve chromatography on an AKTA (GE) purifier. The purity of the protein was checked by SDS-PAGE. After the electrophoretic pure (>95%) protein was obtained, it was concentrated to a higher concentration with an ultrafiltration concentrator (Millipore, USA). Crystal growth w...

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Abstract

The invention discloses a crystal of thymidylate synthetase (ThyX) of helicobacter pylori. The crystal structure is a unit cell that a is equal to 222.15, b is equal to 49.26, c is equal to 142.49, alpha is equal to 90 degrees, beta is equal to 98.65 degrees and gamma is equal to 90 degrees; and the protein space group of the crystal is C2. The crystal structure lays a foundation for designing a small molecular inhibitor and an antibacterial agent.

Description

technical field [0001] The invention belongs to the fields of molecular biology and applied microbiology, and particularly relates to a crystal of thymidylate synthase ThyX of Helicobacter pylori and a preparation method thereof. Background technique [0002] Helicobacter pylori (H. pylori) is a Gram-negative microaerophilic bacterium, which is an important pathogenic factor of chronic gastritis and gastroduodenal ulcer, and is closely related to the occurrence of gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma. The World Health Organization has listed it as a first-class pathogenic factor. According to statistics, the infection rate of H. pylori in the world population is more than 50%, and the multi-drug resistance situation faced by the clinical use of antibiotics to treat H. pylori infection is becoming increasingly severe. Therefore, screening and discovering new drug therapeutic targets has become a major research hotspot in recent years, which h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/00C12N15/52C12N15/63
Inventor 邹全明张晓丽章金勇郭鹰石云毛旭虎郭刚
Owner ARMY MEDICAL UNIV