Transposase-antibody binding protein fusion protein and preparation and application thereof

Abstract

The invention belongs to the technical fields of molecular biology, genomics and biotechnology, and particularly relates to a transposase-antibody binding protein fusion protein and preparation and application thereof. The fusion protein comprises a transposase portion, a linker peptide portion, and a portion capable of binding to an Fc segment of an antibody, and thus, the fusion protein has botha transposition function and an antibody Fc segment binding function. The fusion protein is used to create a new in-vivo protein-chromatin DNA interaction studying method, namely chromatin-transposition-sequencing (ChT-Seq) method. Compared with traditional ChIP-Seq, the ChT-Seq method has the following advantages: fewer steps, high efficiency, saving, good repeatability, less required sample DNA, and greatly-reduced sample and information lost in the process. The method is a powerful tool for studying in-vivo protein-chromatin DNA interaction, and has great significance for research on generegulation and epigenetics and the like.

Description

technical field [0001] The invention belongs to the fields of molecular biology, genomics and biotechnology, and specifically relates to a fusion protein of a transposase-antibody binding protein and its preparation and application. Background technique [0002] Transposon sequences can be inserted and integrated into random positions in the genome under the action of transposases. Because of its random insertion into DNA, transposases are often used in mutant library construction and sequencing library construction. In the library construction link of high-throughput sequencing, transposase can randomly interrupt the sequence to be tested, and add adapters to both ends of the fragmented sequence. After PCR amplification, the next step of sequencing can be directly performed, which is compared with traditional ultrasonic disruption The method has the great advantage that fewer steps save costs and time. [0003] Antibody binding proteins such as ProteinA, ProteinG or Prote...

AI Technical Summary

Problems solved by technology

Similarly, ChIP-Seq inherits the same technical difficulties as ChIP. In addition, in the library construction-sequencing link, the end of the DNA fragment is filled first, then A' is added to the end, and a Y-shaped adapter is added to A', and finally The sequencing method is also cumbersome and has many steps, and each step will lose precious samples and the final sequencing information

Due to the loss, the amount of sample DNA required must also be high, which is a great limitation for some studies that are difficult to obtain a large number of samples, such as some nerve cells that are difficult to culture, some need to study cell heterogeneity, and need to be conducted on a single cell. study tumor cells

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