Study Of Nanometer Carrier Modified Exogenous Gene During Cell Transfection In Vitro

Objective:Nanotechnology is a kind of emerging technology based on nanomaterials.With the small volume property, nanomaterials can penetrate physical barriers andpromote absorb and efficacy of medicine. On the other hand, the half-life ofmedicine can be prolonged with the protection of nanomaterials and the plasmaconcentration can keep for a longer time. Being a kind of economic and efficientapplication, nanomaterials still have many drawbacks，such as low transfectionefficiency. Some of them even have side effect. By comparison with conventionalPEI nanocarriers，this paper aims to explore the transfection efficiency of ZM5K5, anew type nanocarriers which deriving from zein. All nanocarriers carried p-EGFPplasmid to confirm the result of transfection. What we did will provide a new way ofthinking for the applications of nanocarriers in many fields, such as biologicalScience, materials science, chemistry and clinical medicine.Methods:Firstly, Top10, an efficient engineering e. coli strains, which carried p-EGFPplasmid was amplified in LB liquid medium with100mg/ml ampicillin. Then theDNA plasmid was extracted and the concentration was measured. The DNAplasmids were packaged in ZM5K5, a new type of nanometer carrier deriving fromzein, or traditional PEI nanometer carrier. After human lung adenocarcinoma cellsA549were cultured, the ZM5K5or PEI carrying DNA plasmids were transfectedinto the cells. At last, different Serum concentration and plasmid concentration wereset to find the best transfection conditions.Results:1. After amplification in LB liquid medium, the bacteria solution presented milk white and bacteria growth in logarithmic phase. The final concentration ofTop10was measured and the result of OD260/280was0.4. About0.6grams bacteriawas got from bacteria solution after centrifugation. Then DNA plasmids wereextracted and the final concentration was2.63μg/μl.2. Human lung adenocarcinoma cells A549growth well under the condition ofDMEM medium with10%fetal bovine serum. After seeding, the cells adhere welland proliferate rapidly. The transfection was carried out at the point of80%cellconfluence.3. Comparing with traditional PEI nanometer carrier, the new typenanocarriers ZM5K5could transfer DNA plasmids more efficient into A549cells.What was more, the green fluorescence of cells transfected by ZM5K5was morestable.4. With repeated experiments，the best transfection conditions of nanocarrierZM5K5with DNA plasmids were got. The best concentration of fetal bovine serumwas10%and the most appropriate dilution ratio of plasmid DNA was1:8. At last thebest culture condition was37℃and5%CO2for36hours.Conclusion：1. ZM5k5can be used as a new type nanocarriers for cell transfection, and besuccessfully transfected in A549cells.2. DNA plasmids transfection efficiency of A549cells with new typenanocarrier ZM5k5is higher than that of traditional PEI nanocarrier.3. The dosage of the nanoparticle carrier ZM5K5and concentration of DNAdirectly affect the transfection efficiency.