鸡MMP7稳定过表达DF-1细胞系的构建与鉴定

doi:10.11843/j.issn.0366-6964.2025.12.045

Abstract

Abstract: This study aimed to establish a DF-1 cell line stably overexpressing chicken mmp7 gene, providing a foundation for investigating the role of mmp7 in related diseases at the cellular level. The chicken mmp7 gene sequence was synthesized and cloned into the lentiviral vector pLVX-MMP7-IRES-puro. Using a three-plasmid lentiviral packaging system, the recombinant plasmid pLVX-MMP7-puro was co-transfected with the helper plasmids pCMV-VSV-G and psPAX2 into 293T cells at the logarithmic growth phase to produce mmp7-carrying lentiviral particles. DF-1 cells were infected with the recombinant lentivirus for 72 h, followed by puromycin selection (1.5 μg·mL^-1). Integration of mmp7 into the DF-1 genome was confirmed by PCR, while its transcriptional and protein expression levels were assessed via RT-PCR and Western blot, respectively. The CCK-8 assay was used to evaluate the effect of MMP7 overexpression on DF-1 cell proliferation. A DF-1 cell line stably overexpressing chicken MMP7 was successfully generated. Overexpression of MMP7 showed no significant impact on cell viability at 24-48 h but led to a notable reduction at 120-144 h (P<0.05). This cell line serves as a valuable tool for further exploration of MMP7’s biological functions.

Key words: matrix metalloproteinase-7, lentivirus, stable overexpression, DF-1 cell line

CLC Number:

DENG Mengling, SUI Minmin, SONG Haiyang, YANG Jianfa, ZOU Fengcai, HE Junjun. Construction and Identification of a Chicken MMP7 Stable Overexpression DF-1 Cell Line[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6442-6449.

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Construction and Identification of a Chicken MMP7 Stable Overexpression DF-1 Cell Line

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