A kind of cultivation method for transfecting human tumor suppressor gene green fluorescent protein carrier

Abstract

A method for the cultivation of human tumor suppressor gene green fluorescent protein carrier, design primers on GENBANK with reference to human anti-cancer gene sequence, collect peripheral blood of early lung cancer patients to extract total RNA, and use reverse transcription polymerase chain reaction (RT-PCR) Amplify the P53 fragment, clone the gene fragment directionally onto the pMD18-T vector, construct the pMD18-P53 recombinant plasmid, identify the recombinant plasmid by PCR amplification, enzyme digestion and sequencing, and digest the pMD18-P53 Identify and extract the directional clone of the P53 target gene into the pEGFP-N1 eukaryotic expression vector to construct the pEGFP-N1-P53 expression vector. The beneficial effects of the present invention are: using the eukaryotic expression vector pEGFP-N1, which is safe, stable, strong in replication, high in expression rate, and easy to detect, as the transport carrier of human anti-cancer genes, thereby constructing a safe, stable, easy-to-detect, safe and reliable The eukaryotic expression vector pEGFP-N1-P53 is used for the prevention and treatment of cancer.

Description

technical field [0001] The invention relates to a method for cultivating a carrier, in particular to a method for cultivating a human tumor suppressor gene green fluorescent protein carrier, pEGFP-N1-P53. Background technique [0002] With the improvement of human living standards, serious air and water pollution, pesticide residues and other indirect effects on human health, cancer is now recognized as one of the deadly killers. According to the report of the International Cancer Research Center (IARC), in 2002, there were 10.9 million new cases of cancer worldwide, 6.7 million deaths, and 24.6 million current cases. The number of cancer cases is increasing at an average annual rate of 3%-5%, and the number of cases and deaths has increased by 24.7% and 19.2% respectively compared with 10 years ago. It is estimated that by 2020, there will be 20 million new cases and 12 million deaths worldwide. The treatment of cancer is still a worldwide problem. A single surgical treat...

AI Technical Summary

Problems solved by technology

The treatment of cancer is still a worldwide problem. A single surgical treatment is likely to cause the metastasis of the lesion, while radiotherapy and chemotherapy bring great pain to the patient and at the same time inhibit the bone marrow and cause pancytopenia; red blood cell reduction causes anemia, leukopenia causes infection, platelet Reduce bleeding, any of which could be life-threatening

Modern medicine tries to use gene therapy to prevent and treat cancer. However, there are still many theoretical and technical problems that are difficult to overcome for transgenes. The vectors are difficult to express in animals, have low safety, weak replication, and are difficult to detect.

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