Opened protein diversity display method

Abstract

The invention relates to a method for analyzing protein expression difference directly on protein level. The invention has the technical proposals as follows: a fit DNA or RNA random sequence library is constructed, and the cell protein in the DNA or RNA aptamer random sequence library is extracted, so as to obtain a DNA or RNA random sequence library that can reflect protein level change; after subtractive hybridization of target cells and control cells in the DNA or RNA random sequence library that can reflect protein level change is carried out, PCR clone is carried out to eliminate sequence, so as to obtain a differential display library, and sequencing is carried out; the corresponding type of DNA or RNA aptamer is prepared by amplification of marked primer; and the DNA or RNA aptamer is filtered to determine specific proteins having expression difference and function difference. The method has differential display that transfers the expression difference on protein level onto DAN or RAN level by aptamer, thereby achieving the purpose of simply and quickly filtering and expressing differential proteins in high throughput.

Description

technical field [0001] The invention relates to a biological protein analysis method, in particular to a method for directly analyzing protein expression differences at the protein level. Background technique [0002] Before the present invention, there are currently two types of differential analysis methods for protein expression, one is the differential analysis method at the RNA level, and the other is the differential analysis method at the protein level. There are two main types of the former, one is the differential display PCR method (DD-PCR for short), and the other is the subtractive hybridization method and suppression subtractive hybridization method (SSH for short). [0003] The main steps of the DD-PCR method are to first extract the mRNA of the tissue sample, and then perform reverse transcription reaction with 12 pairs of anchor primers at the 3′ end; perform PCR amplification with the anchor primers and 20 random primers at the 5′ end, and perform PCR amplif...

AI Technical Summary

Problems solved by technology

However, due to the difficulty of protein manipulation, the research on the difference display at the protein level is now mostly using the antibody chip method, which can only find the expression difference changes of known proteins and cannot find new proteins, and the current cost of antibody chips Extremely high, its use requires precision instruments, ordinary scientific research institutes cannot afford it, and the number of antibodies contained in antibody chips is limited, which is far less than the number of gene chips, and cannot really achieve the purpose of high-throughput screening

[0004] On the other hand, the existing technology also provides an open differential display analysis method at the protein level, such as a large-scale instrument for two-dimensional electrophoresis plus mass spectrometry, but the price is high and it is not suitable for small and medium-sized laboratories