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Host cell for expressing KCNQ1/KCNE1 protein and preparation method of host cell

A KCNE1, host cell technology, applied in the field of host cells expressing channel proteins and their preparation methods, can solve the problems of no observed cytotoxicity, rarely seen cytotoxicity, and little, etc., achieve good biological safety, improve The amount of drug screening, the effect of simple preparation

Active Publication Date: 2015-07-01
WUXI BIOLOGICS CO LTD +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] (1) Many cell lines and primary cells can be efficiently transduced, but little or no cytotoxicity is observed, even at MOIs as high as 10,000

Method used

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  • Host cell for expressing KCNQ1/KCNE1 protein and preparation method of host cell
  • Host cell for expressing KCNQ1/KCNE1 protein and preparation method of host cell
  • Host cell for expressing KCNQ1/KCNE1 protein and preparation method of host cell

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Embodiment 1

[0054] 1. Construction of recombinant baculovirus vector

[0055] According to the full gene sequence of KCNQ1 [Homo sapiens potassium voltage-gated channel, KQT-like subfamily, member 1 (KCNQ1), NCBI Reference Sequence: NM_000218.2, the full gene sequence is shown in SEQ ID NO.1) and the full gene sequence of KCNE1 [ Homo sapiens potassium voltage-gated channel, Isk-related family, member 1 (KCNE1), NCBI Reference Sequence: NM_000219.3, full gene sequence as shown in SEQ ID NO.2], design its specific primers, wherein:

[0056] The upstream primer sequence of KCNQ 1: 5'-ACTGAATTCGCCACCATGGCCGCGGCCTCCTC-3', as shown in SEQ ID NO.3, contains an EcoRI restriction site;

[0057] The downstream primer sequence of KCNQ 1: 5'-CTGAAGCTTCAGGACCCCCTCATCGGGG-3', as shown in SEQ ID NO.4, contains a HindIII restriction site;

[0058] The upstream primer sequence of KCNE 1: 5'-ACTGAATTCGCCACCATGATCCTGTCTAACACCACAGCG G-3', as shown in SEQID NO.5, contains an EcoRI restriction site;

[0059...

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Abstract

The invention discloses a host cell for expressing KCNQ1 / KCNE1 protein and a preparation method of the host cell. The hose cell is formed by the construction of a BacMam recombinant baculovirus vector, preparation of the BacMam recombinant baculovirus vector and after the BacMam recombinant baculovirus vector is transformed into CHO (Chinese Hamster Ovary) cells. Through the host cell, KCNQ1 / KCNE1 channel protein can be obtained, and a research basis is provided for high-throughput drug screening.

Description

technical field [0001] The invention relates to a host cell expressing a channel protein and a preparation method thereof, in particular to a host cell expressing a KCNQ1 / KCNE1 channel protein using a BacMam virus and a preparation method thereof. Background technique [0002] The KCNQ potassium channel is an important branch of the potassium channel superfamily. The potassium channel encoded by the KCNQ gene is distributed in the ganglion, cerebral cortex, hippocampal nerve cells, cardiomyocytes, inner ear hair cells, etc., and plays an important role in regulating nerve excitability and myocardial action potential. and auditory aspects. KCNQ is divided into five subtypes, namely KCNQ1, KCNQ2, KCNQ3, KCNQ4, and KCNQ5. Among them, KCNQ1, as a member of the potassium channel family, plays an important role in the physiological regulation and pathological mechanism of the heart, inner ear, and small intestine. [0003] KCNQ1 (KvLQT1) was first discovered at a chromosomal locu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/866C12N15/66C12N5/10
Inventor 楼庄伟董海恒宋云鹏赵娜陈英梅佳贾园陈侃孔云华吴瑶吕强
Owner WUXI BIOLOGICS CO LTD
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