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A fusion protein containing apce2 mutant and its application

A fusion protein and cell technology, applied in the biological field, can solve problems such as weak fluorescence and achieve the effect of improving brightness

Active Publication Date: 2021-11-30
HUAZHONG AGRI UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A fusion protein containing apce2 mutant and its application
  • A fusion protein containing apce2 mutant and its application
  • A fusion protein containing apce2 mutant and its application

Examples

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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 a fusion protein, which is characterized in that: the amino acid sequence of the fusion protein is: F1 polypeptide—connection protection peptide 1—F2 polypeptide—connection protection peptide 2—F1 polypeptide, wherein the sequence of F1 polypeptide is SEQ ID No .1 and any one of SEQ ID No.2, and the F2 polypeptide sequence is any one of SEQ ID No.3 to SEQ ID No.14. In the present invention, a fusion protein is obtained by fusing the ApcE2 mutant with the genetically engineered allophycocyanin subunit BDFP1.1 or BDFP1.2, which improves the weak fluorescence of the ApcE2 mutant in mammalian cells The problem. It is also found that adding phytochrome PΦB in vitro during the fluorescent labeling process can effectively improve the brightness of the fusion protein for cell labeling.

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 Patents(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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