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The G2019S, K1906M, R1441C, N1437S and T1348N mutation were introduced by site-directed mutagenesis (QuikChange™ XL Site-Directed Mutagenesis Kit, Stratagene, La Jolla, USA).
First, the A159-K160 deletion and mutation were introduced into a full-length IBV infectious clone, and the effect of this minimal deletion and mutation on the replication of viral RNA and the recovery of infectious IBV was analyzed.
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The T315I mutation was introduced by site-directed mutagenesis.
Subsequently, a R419L mutation was introduced into purA to reduce the flux from IMP to AMP.
In brief, the S89G mutation was introduced in exon 6 using an overlap extension-PCR method.
The R217Q mutation was introduced by site-directed mutagenesis.
The AXXLL mutation was introduced by a fusion PCR approach.
The point mutation was introduced with the QuickChange XL Site-Directed Mutagenesis kit (Stratagene).
DNA sequencing was performed to confirm that no undesired mutation was introduced by PCR.
The D174H mutation was introduced by PCR site-directed mutagenesis as described previously [55].
The R217Q mutation was introduced in VSFP2A by site-directed mutagenesis.
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