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Nano cationic liposome carrying FL gene recombinant plastid, and preparation method and application of nano cationic liposome

A cationic liposome and recombinant plasmid technology, applied in the field of biomedicine, can solve the problems of low transfection efficiency, unsatisfactory target distribution, and poor stability, and achieve simple preparation methods, good application prospects, and low immunogenicity Effect

Inactive Publication Date: 2013-01-16
孙念峰
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  • Claims
  • Application Information

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Problems solved by technology

Although cationic liposomes have the advantages of low toxicity, low immune response, and strong modifiability, the transfection efficiency is still low, the target distribution is not ideal, and the stability is poor

Method used

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  • Nano cationic liposome carrying FL gene recombinant plastid, and preparation method and application of nano cationic liposome
  • Nano cationic liposome carrying FL gene recombinant plastid, and preparation method and application of nano cationic liposome
  • Nano cationic liposome carrying FL gene recombinant plastid, and preparation method and application of nano cationic liposome

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Experimental program
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Embodiment Construction

[0028] 1. Gene synthesis

[0029] 1. FLT3L (ie FL) gene synthesis: FLT3L gene sequence was synthesized, and the gene was cloned between Sal Ⅰ and BamH Ⅰ of the puc57 vector. The full length of the FLT3L gene is 510bp, encoding a total of 169 amino acid residues. The synthesis of the FLT3L gene was completed by Shanghai Sangon Bioengineering Technology Co., Ltd.

[0030] 2. Verification of synthetic genes

[0031] The recombinant plasmid puc57-FLT3L cloned with the FLT3L gene was digested and verified, and its sequence was determined to detect whether its nucleotide sequence was completely consistent with the designed one (the determination results were as follows: figure 2 shown). Sequence determination was completed by Shanghai Yingjun Company.

[0032] 3. DNA digestion

[0033] (1) Number the clean, dry and sterilized Eppendorf tubes, add 3 μL of the above-mentioned plasmid and 1.5 μL of 10×Taq Buffer with a micropipette, then add redistilled water to make the total volu...

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Abstract

The invention discloses a nano cationic liposome carrying an FL gene recombinant plastid, and a preparation method and application of the nano cationic liposome, and belongs to the field of biological medicine. The liposome comprises Lipofectamine 2000 and a recombinant plasmid pEGFP-cl-FL carrying an FL gene according to an electrostatic adsorption principle, wherein the recombinant plasmid pEGFP-cl-FL carrying the FL gene is prepared by cloning the FL gene into a pEGFP-cl vector containing a green fluorescent protein reporter gene. The nano cationic liposome has the characteristics of simple preparation method, low toxicity, low immunogenicity, slow release and high-efficiency targeting, is particularly suitable for serving as a gene vector of in vivo and in vitro transfection and gene therapy, and has a good application prospect in the gene research aspect in the fields of tumor and others.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a novel nano cationic liposome capable of efficiently carrying FL gene recombinant plasmids, a preparation method and an application. Background technique [0002] Colon cancer is one of the most common digestive tract malignancies, and its morbidity and mortality are increasing year by year, seriously threatening human health. Surgical treatment is one of the traditional treatment methods. Although it can remove tumor lesions, it cannot completely remove all cancer cells in the body, and the remaining cancer cells can cause tumor recurrence and metastasis. Radiotherapy and chemotherapy, as powerful adjuvant treatments to surgical treatment, are of great benefit in eliminating residual cancer cells and preventing tumor recurrence and metastasis. However, due to their serious side effects and the possible insensitivity or Drug resistance has greatly limited its clinical appl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63A61K48/00A61K9/127A61P35/00
Inventor 孙念峰田爱玲胡三元陈景波黄庆先
Owner 孙念峰
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