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Fusion protein containing ApcE2 mutant and application of fusion protein containing ApcE2 mutant

A fusion protein, 1-F2 technology, applied in the biological field, can solve the problems of weak fluorescence and achieve the effect of improving brightness

Active Publication Date: 2019-04-02
HUAZHONG AGRI UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] After the applicant used ApcE2 (24-245) as a template for genetic modification in the research, the BDFP3 and other series of proteins obtained solved the solubility problem. After combining with the phytochrome PΦB, E. coli cells can be fluorescently labeled, but in mammals The fluorescence of the label in the cell is weak

Method used

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  • Fusion protein containing ApcE2 mutant and application of fusion protein containing ApcE2 mutant

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Embodiment 1

[0048] The ApcE2 mutants used in this example are BDFP3 and BDFP3(Δ88-106), and the amino acid sequences are shown in SEQ ID No.9 and SEQ ID No.11. figure 1 It is the overlapping diagram of the fluorescence emission spectrum of BV-BDFP1.1 / 1.2 and the absorption spectrum of PΦB-BDFP3. It can be seen that the absorption fluorescence spectra of BDFP1.1 and BDFP1.2 used for fusion overlap, which is beneficial to fluorescence energy transfer and can Efficiently transfer energy to BDFP3 series proteins.

[0049] A fusion protein BDFP1.1:3:1.1, the amino acid sequence of which is shown in SEQ ID No.17.

[0050] A fusion protein BDFP1.2:3:1.2, the amino acid sequence of which is shown in SEQ ID No.18.

[0051] A fusion protein BDFP1.2:3(Δ88-106):1.2, the amino acid sequence of which is shown in SEQ ID No.19.

[0052]

[0053]

[0054] The gene sequence encoding the above fusion protein was cloned into Escherichia coli, and expressed and recombined with phytochrome PΦB in vivo ...

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Abstract

The invention discloses fusion protein. The fusion protein is characterized in that the amino acid sequence of the fusion protein is F1 polypeptide, connected protective peptide 1, F2 polypeptide, connected protective peptide 2 and F1 polypeptide, wherein the sequence of the F1 polypeptide is any of SEQID No.1 and SEQID No.2; and the sequence of the F2 polypeptide is any of SEQID No.3 to SEQ ID No.14. An ApcE2 mutant and allophycocyanin subunits BDFP1.1 or BDFP1.2 which are subjected to genetic engineering are merged to obtain the fusion protein, and the problem that the ApcE2 mutant marked fluorescence in lactation animal cells is weak, is solved. In the fluorescence marking process, phytochrome P phi B is added in vitro, so that the brightness of the fusion protein for cell labelling canbe effectively improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fusion protein containing an ApcE2 mutant and its application. Background technique [0002] The development of modern biology relies more and more on optical technologies, such as fluorescence imaging, optical detection and light-induced manipulation, etc. For imaging and positioning of deep tissues in vivo, it is best to use fluorescence in the 650nm-900nm band. Because the light in this band has less autofluorescence, less light scattering and less damage to tissues. In recent years, two types of proteins that have been studied more as fluorescent probes are the green fluorescent protein family and bacterial phytochrome protein (BphP). [0003] In order to better adapt to the environment and capture more energy from the environment, a small number of cyanobacteria slowly evolved phycobilisomes that can utilize far-red light. In these phycobilisomes, there are usually differen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00G01N21/64G01N33/68G01N33/58
CPCC07K14/195C07K14/795C07K2319/00G01N21/64G01N33/582G01N33/68
Inventor 周明夏坤付卫雷吴明陈彦蓉
Owner HUAZHONG AGRI UNIV
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