Differential proteomic analysis method using transcriptome sequencing and based on combination of iTRAQ and LC-MALDI

Abstract

The invention discloses a differential proteomic analysis method using transcriptome sequencing and based on combination of iTRAQ and LC-MALDI. The method comprises the steps: (1) transcriptome sequencing is performed; (2) differential proteomic analysis is used for extracting the total protein fluid of livers of normal group rats and liver cancer group rats, the two groups are mixed respectively,proteins are precipitated and quantified by an acetone precipitation method, each group contains about 100 [mu]g of the total proteins, trypsin enzymolysis, reduction alkylation, iTRAQ labeling and the like are performed successively, and the process is performed by strictly following instructions of an iTRAQ kit; and (3) bioinformatics analysis is performed. A clinical sample is adopted for positioning and quantitative expression verification, an evidence of the clinical sample with clinical relevance is searched, clinical value is evaluated, and a new clue is provided for research of livercancer pathogenesis and a liver cancer mechanism. Liver cancer key molecules screened by the project can lay the research basis for exploration of liver cancer markers related to early detection, classification and prognosis evaluation, and selection of more effective and accurate liver cancer treatment target positions.

Description

technical field [0001] The invention relates to the fields of gene transcriptomics and proteomics, in particular to a transcriptome sequencing and differential proteomics analysis method based on iTRAQ combined with LC-MALDI. Background technique [0002] One of the most important fields in biological and medical research is the observation of the structure, function and regulation of biological systems and life processes. However, in the past few hundred years, biologists have been focusing on the expression changes and functions of individual genes or proteins in biological systems, but cannot study the changes in living systems from a global and holistic perspective. With the advancement of medicine, people have found that many diseases, especially cancer, are often caused by the synergistic effects of multiple factors, multiple genes, and multiple pathways. This requires a technology and means that can comprehensively, dynamically and systematically study the life syste...

AI Technical Summary

Problems solved by technology

However, with the completion of the Human Genome Project, it was found that only from the perspective of genomics can not completely correctly predict the splicing, splicing, and the start and end positions of open reading frame codons during translation and post-translation various modifications of