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Mutational analysis identifies leucine-rich repeat insertions crucial for pigeon toll-like receptor 7 recognition and signaling.

Vet Immunol Immunopathol. 2015; 
Xiong D, Song L, Jiao Y, Kang X, Chen X, Geng S, Pan Z, Jiao X.
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Custom Vector Construction … expression vector pCMV-HA (Invitrogen, Carlsbad, CA, USA) digested with SalI and KpnI (Takara Biotechnology Co., Dalian, China) to create pCMV-PiTLR7DelxINy. The sequences of the resulting constructs were confirmed with DNA sequencing (Genscript, Nanjing, China) … Get A Quote

摘要

Toll-like receptor 7 (TLR7) is responsible for recognizing viral single-stranded RNA and antiviral imidazoquinoline compounds, leading to the activation of the innate immune response. In this study, mutated pigeon TLR7 fragments, in which the insertion at position 10 of leucine-rich repeat 10 (LRR10) or at position 15 of LRR2/11/13/14 was deleted, were amplified with an overlap-PCR method, and inserted into the expression vector pCMV. The immune functions of the TLR7 mutants were determined with an NF-κB luciferase assay of transfected cells. The deletion of the insertions absolutely abolished TLR7-NF-κB signaling. With quantitative real-time PCR and sandwich enzyme-linked immunosorbent assay, we observed tha... More

关键词

Innate immunity; LRR insertion; Mutational analysis; Pigeon; TLR7