Figure 3.

Viral clone isolation from plaques and analysis of amino acid mutations. (A) Schematic diagram of amino acid mutations in the S protein of parental SARS-CoV-2. The mutations were analyzed by whole-genome sequencing of SARS-CoV-2 virus isolated from the tumor #7 at 30 dpi as shown in Supplementary Table 3 . (B) Analysis of amino acid mutations in the S protein of parental SARS-CoV-2 and its variants in lung tumor. cDNA was synthesized from the pool of viruses in the supernatants of SARS-CoV-2 parental virus-infected tumor homogenates obtained at 30 dpi. The sequences of the S1 subunit region of the S protein were cloned and analyzed by direct DNA sequencing. (C) Viral clone isolation from plaques and analysis of amino acid mutations. Viral isolates were isolated from plaques derived from supernatants of the tumor infected with parental SARS-CoV-2 at 30 dpi. Fifteen viral isolates collected and then cDNA was synthesized from each clone. The S1 subunit region of the S protein was further cloned and analyzed by direct DNA sequencing. Three selected viral isolates (S-1, S-6, S-9) were collected, and their whole-genome sequences were analyzed. Each panel indicates the accumulation and substitutions of mutations at an amino acid position in parental SARS-CoV-2. Origin (Parent): amino acid sequences of parental SARS-CoV-2 ( Supplementary Table 1 ). dpi, days post-infection.