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Objective: investigating the influence of the hepatitis C virus F protein deletion on viral replication and pathogenic infection and the results were analyzed. Methods: the experimental method selected biochemical methods, the first step, the preparation of the hepatitis C virus RNA transcript; the second step, according to the result of the correlation detection culture experiments, including: PCR technology, the detection of infectious replication, immunofluorescence, transfection, etc. Finally, the relevant results to collate, record and analyze. Results: the introduction of the mutant gene expression and the number of the original number of genes expression was no significant difference in the expression levels of the two groups and the core protein also no significant difference (P <0.05); F protein expression in tumor tissue than other tissues, indicating HCC formation and F protein has some relevance;Core protein and the wild-type core protein contains a single mutation in the structure remained the same; F-deficient secondary structure of the core protein has more variations. Conclusion: HCV F translation and replication process of the missing protein of the virus had no effect on viral translation, reproduction and medical treatment to increase the resistance of the secondary structure of the core protein. Introduction Hepatitis C virus infection is the culprit leading to chronic liver disease and the resulting infection can be further developed into cirrhosis, fatty liver and liver cancer and other serious diseases [1]. There are about 170 million carriers of hepatitis C virus worldwide, and about 100,000 people have developed cancer patients. It is generally believed that the route of transmission of hepatitis C virus includes blood and blood products, the current drug for hepatitis C virus treatment only interferon and the drug's response rate is very low, side effects are more expensive, does not meet the requirements, and Hepatitis C virus vaccine is still in the research stage [3]. The hepatitis C virus genome consists of two parts, which are located in the middle of the open reading frame and at both ends of the non-coding region (conservative), the length can reach 9600bp. The noncoding region is indispensable for the viral RNA replication process and the translation process [4-5]. A single open reading frame translates as many as 3,000 amino acid residues, which are polyprotein precursors, which can be cleaved into six nonstructural proteins and 10 species under the dual action of viral proteases and host cells Structural proteins (E1, E2, C and P7). This year, some laboratories have found a viral protein called the reading frame shift protein, also known as core + 1 protein. In addition, there are also found that the endogenous codon can also produce F protein, the starting site is mainly located in the 85/87 codon was named core + 1 / S [6]. From the discovery of F protein to now has more than ten years of history, but the human biology of its red to know little about how to understand the hepatitis C virus F protein is how the virus infection, disease and replication process, the study observed the process of viral translation and replication in the absence of F protein, and further analyzes how the secondary structure of the core protein changes after its deletion. Materials and Methods Reagents and Instruments. The name of the reagent and the instrument and the details of the 2017 International Conference on Materials Science and Biological Engineering (ICMSBE 2017) Copyright © (2017) Francis Academic Press , UK 33 manufacturer are shown in Table 1. Table 1 Reagent and instrument name and manufacturer Reagent or instrument name Production company or individual QuikChang LinghrningSite-Directed Mutagenesis Kit Stratagene Corporation (USA) products Isopropyl thiogalactoside Beijing tripod biotechnology limited liability company 5-bromo-4-chloro-3-indole-β-D galactoside Beijing tripod biotechnology limited liability company NZ amines Beijing tripod biotechnology limited liability company Yeast extract Beijing tripod biotechnology limited liability company 14ml BD Falcon Polypropylene Tube Shanghai Ya Yi Science Experimental Equipment Co., Ltd HCV NS5A monoclonal antibody Charles M .Riceprofessor T7 in vitro transcription kit Promrga company M-MLV reverse transcriptase Promrga company Fetal bovine serum America invitorgen company DMEM America invitorgen company Opti-MEM America invitorgen company Alex488 goat anti-mouse fluorescent America invitorgen company FITC-goat anti-human IgG America invitorgen company Lipofectamine 2000 America invitorgen company RNA extraction reagent Shanghai Huashun Biological Engineering Company Construction of Cell Lines and Plasmids. The introduction of the kit was carried out in the presence of PFL-J6JFH1 as template, and the number of introduced mutations was 5, and the purpose of introducing these mutations was to terminate the replication of all known F proteins, and this process did not Change the amino acid sequence of hepatitis C virus. Preparation of Viral RNA Transcripts. In the first step, the linearization treatment: the plasmid is linearized by XbaI; the second step, in vitro transcription: the preparation of RNA in vitro transcripts by T7 transcription kit; the third step, purity and integrity detection: reagent A mixture of phenol, isoamyl alcohol and chloroform at a volume ratio of 25: 1: 24 was selected and the purity and integrity of the mixture were tested by electrophoresis with 1.5% agarose gel. RNA Transcript Transfection and Cell Culture. Transfection of RNA transcripts: 1000 g of trypsin digested cells were washed and centrifuged, separated, washed, resuspended, diluted, mixed and clicked (270V, 100mΩ) and then transferred to a constant temperature incubator (5% CO2, 37 ° C). Cell culture: The culture medium, additives and conditions required for transfection are shown in Table 2.

Impact Analysis of the Hepatitis C Virus F Protein Deletion on Viral Replication and Pathogenic Infection