鸡marco稳转巨噬细胞系的构建与鉴定

doi:10.11843/j.issn.0366-6964.2024.11.046

Abstract

Abstract:

The study aimed to establish the HD11 cell line with a stable expression of the marco gene through the lentivirus expression system, providing a crucial tool for investigating the antiviral immune function of the marco gene. Initially, the marco gene coding sequence was amplified by PCR using HD11 cells as a template and cloned into the lentiviral vector to generate the recombinant lentiviral plasmid pLV3-CMV-MCS-Puro-marco. Subsequently, the recombinant plasmid pLV3-CMV-MCS-Puro-marco was co-transfected with helper plasmids pLP1, pLP2, and pMD2.G into 293T cells using a tetra-plasmid lentivirus packaging system, resulting in the construction of recombinant lentivirus carrying the chicken marco gene. The successfully packaged recombinant lentivirus was then used to infect HD11 cells, followed by puromycin (2 μg·mL^-1) selection for 72 h. The stable expression of the chicken marco gene in the HD11 cell line was confirmed by RT-qPCR analysis. This study successfully established a stable HD11 cell line expressing the chicken marco gene, providing a crucial basis for further exploration of the marco gene's function.

Key words: marco cell line, stable expression, lentivirus

CLC Number:

S831.2

Wang GUO, Guangzhong PENG, Hongao HUANG, Huixian WU, Xuming HU, Yu ZHANG, Yang ZHANG, Guohong CHEN, Qi XU. Construction and Identification of the Chicken marco-stable Macrophage Cell Line[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5310-5316.

Figures/Tables 4

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Construction and Identification of the Chicken marco-stable Macrophage Cell Line

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