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LH0147 gene, and expression vector, expression product and application thereof

A technology of LH0147, expression vector, applied in the direction of using vector to introduce foreign genetic material, application, genetic engineering, etc.

Inactive Publication Date: 2020-12-01
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the successful prokaryotic expression of bovine Pasteurella multocida type A adhesin LH0147 protein

Method used

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  • LH0147 gene, and expression vector, expression product and application thereof
  • LH0147 gene, and expression vector, expression product and application thereof
  • LH0147 gene, and expression vector, expression product and application thereof

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Embodiment Construction

[0021] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0022] The sources of reagents and materials in the embodiments of the present invention are as follows: plasmid pET-28a (+) is provided by Novagen; DNA gel recovery kit is provided by Tiangen Biochemical Technology (Beijing) Co., Ltd.; plasmid extraction kit is provided by Tiangen Root Biochemical Technology (Beijing) Co., Ltd.; ClonExpress II One Step Cloning Kit provided by Vazyme; DNA polymerase provided by TaKaRa.

[0023] 1. Primer design and acquisition of target fragments

[0024] According to the coding gene sequence of bovine Pasteurella multocida type A adhesin LH0147, the upstream prim...

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Abstract

The invention discloses a bovine-derived serotype A pasteurella multocida adhesin LH0147 gene as well as an expression vector, an expression product and application of the bovine-derived pasteurella multocida adhesin LH0147 gene. The LH0147 gene is shown as SEQ ID NO.1; the amino acid sequence of the LH0147 protein encoded by the LH0147 gene is shown as SEQ ID NO. 2; the recombinant expression vector of the LH0147 gene can be used for expressing the LH0147 protein in vitro; the specific method comprises the following steps: performing cloning to obtain the LH0147 gene according to claim 1, andinserting the LH0147 gene between an enzyme cutting site salI and an EcoRI of an expression plasmid pET-28a to obtain a recombinant expression vector pET-28a-LH0147; converting the recombinant expression vector ET-28a-LH0147 into a competent cell BL21 (DE3) through heat shock, and performing screening to obtain a recombinant genetically engineered bacterium; inoculating the recombinant genetically engineered bacterium in vitro, and inducing the recombinant genetically engineered bacterium to express the LH0147 protein. According to the invention, the gene sequence of the bovine-derived serotype A pasteurella multocida adhesin LH0147 is expressed in vitro for the first time, so that the function and pathogenesis of the protein can be researched.

Description

technical field [0001] The invention belongs to the field of genetic bioengineering, and in particular relates to a bovine-derived Pasteurella multocida type A adhesin LH0147 gene, an expression vector, an expression product and application thereof. Background technique [0002] Pasteurella multocida type A (Pasteurella multocida, Pm) is a Gram-negative bacterium that can cause a variety of pasteurellosis in livestock and poultry, including fowl cholera, bovine hemorrhagic sepsis, and swine pneumonia. This bacterium is widely distributed all over the world and can be transmitted among animals. Animal bites can also cause human infection. The natural medium of transmission is mainly ticks and fleas. Pasteurella multocida is a normal flora of the upper respiratory tract of healthy dairy cows and is a commensal bacterium of the upper respiratory tract. It is an opportunistic pathogen. When animals are affected by certain factors such as long-distance transportation, crowding, ...

Claims

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Application Information

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IPC IPC(8): C07K14/285C12N15/70C12N15/31C12N1/21C12R1/19
CPCC07K14/285C12N15/70
Inventor 周振雷潘子豪程艳
Owner NANJING AGRICULTURAL UNIVERSITY
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