Hepatitis B virus antisense RNA expression vector targeting liver cancer and its construction process and use
A technology of hepatitis B virus and expression vector, applied in virus/bacteriophage, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of lack of specificity in transfection, difficult to achieve gene therapy, etc., and achieve good inhibitory effect and good inhibitory effect Effect
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[0164] 1. Plasmids and strains: Plasmid pBR322-HBV is preserved in our laboratory, which contains 3.2kb HBV DNA of 2 copies of ayw subtype connected head to tail; vector pEBAF is provided by Dr. Yan Ziying, Institute of Virology, Chinese Academy of Preventive Medicine, with a total length of 15.5 Kb. Escherichia coli JM109 and DH5α are the strains preserved in this laboratory.
[0165] 2. Cell line and culture medium: The liver cancer cell line HepG2.2.15 integrates the HBV genome, can transcribe and translate HBV genes, and produce HBsAg, HBeAg, and Dane’s particles. The cell line is quoted from the Institute of Immunology, Shandong Academy of Medical Sciences. The human liver cancer cell line BEL7402 was passaged in this laboratory, expressing alpha-fetoprotein 200ng / ml. Human umbilical vein endothelial cells ECV-304 were donated by the Military Medical College, and the secretion of alpha-fetoprotein was not detected.
[0166] 3. Tool enzymes: Xbal, T4DN...
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