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Preparation of anti-PML protein nuclear location signal antibody and application thereof in APL diagnosis

A technology for locating signals and protein cores, which can be used in anti-animal/human immunoglobulins, material testing products, measuring devices, etc., and can solve problems such as indistinguishability

Active Publication Date: 2014-09-10
AFFILIATED YONGCHUAN HOSPITAL OF CHONGQING MEDICAL UNIV
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Problems solved by technology

Currently, synthetic PML protein N-terminal or RBCC domain peptides are generally used as immunogens in the world, and rabbits or mice are immunized to prepare specific antibodies. The antibodies obtained in this way can not only specifically detect wild-type PML proteins, but also Can detect PML (NLS-) protein, cannot distinguish between the two

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  • Preparation of anti-PML protein nuclear location signal antibody and application thereof in APL diagnosis
  • Preparation of anti-PML protein nuclear location signal antibody and application thereof in APL diagnosis
  • Preparation of anti-PML protein nuclear location signal antibody and application thereof in APL diagnosis

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Embodiment Construction

[0037] Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. The experimental methods not indicating specific conditions in the preferred embodiments are carried out according to conventional conditions.

[0038] Material

[0039] 1.1 Plasmids, strains and animals

[0040] E. coli BL21 bacteria were preserved by the Clinical Diagnostic Laboratory of Chongqing Medical University.

[0041] Construction of the prokaryotic expression plasmid PET-His-NLS-RARα: using the pCMV-HA-NLS-RARα plasmid as a template, respectively using p1:5’-TTA GGATCC ACAGTCAGTGCCCGGGC-3' and p2:5'-GCC AAGCTT TCACGGGGAGTGGGTGGC-3' is the upstream and downstream primers, PCR amplifies the NLS-RARα gene and introduces BamH and Hind restriction sites (underlined parts) at both ends, and inserts it into the prokaryotic expression vector pET-his to construct the prokaryotic expression plasmid The construction of pET-his-NLS-RARα w...

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Abstract

The invention relates to preparation of an anti-PML protein nuclear location signal antibody and application thereof in APL diagnosis. The antibody is prepared from an amino acid sequence shown in SEQ ID NO:1, namely NLS polypeptide as immunogen. The antibody can be used for distinguishing the PML protein from the PML protein lack of a nuclear localization signal, and can be applied to preparation of a diagnostic agent for acute promyelocytic leukemia. In addition, the invention also provides a location method of the PML protein. Therefore, the foundation is established for further research of the PML for inducing cell apoptosis on the gene transcription level to inhibit tumor growth. Meanwhile, wild PML protein and PML (NLS-) protein are differentially detected, and a novel basis is provided for clinical diagnosis of APL and intensive study of pathogenesis of the APL.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the preparation of an anti-PML protein nuclear localization signal (NLS) antibody and its application in APL diagnosis. Background technique [0002] The occurrence of acute promyelocytic leukemia (APL) is closely related to chromosomal translocation, and more than 95% of APL are characterized by chromosomal t(15,17) translocation, forming and expressing PML-RARα fusion protein. Studies have found that the PML-RARα fusion protein can be cleaved by neutrophil elastase (NE) into two variant proteins of about 52KDa ​​and 61KDa. Due to the specific cleavage of NE enzyme, a mutant PML protein lacking nuclear localization signal (NLS) is produced, namely PML(NLS-) protein and NLS-RARα protein with nuclear localization signal . [0003] As a tumor growth suppressor, PML protein has a certain inhibitory effect on the occurrence and development of tumors of different tis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18G01N33/68G01N33/574
Inventor 刘北忠阳小群蒋开玲钟梁
Owner AFFILIATED YONGCHUAN HOSPITAL OF CHONGQING MEDICAL UNIV
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