Bioscience Methods 2024, Vol.15, No.5, 216-225 http://bioscipublisher.com/index.php/bm 220 Figure 2 Mutations in 28 PgABCA2 cDNA sequences from five Cry2Ab-resistant pink bollworm larvae from the CRISPR-R2 strain that lacked gDNA mutations in sgRNA target sites (Adopted from Fabrick et al., 2021) Image caption: (A) The predicted PgABCA2 protein includes amino (N) and carboxyl (C) termini and transmembrane domains (TMD1 and TMD2). Each TMD contains six transmembrane-spanning regions (TM, green), three extracellular loops (ECL, purple), and two intracellular loops (ICL, blue). The two TMDs are connected by a single intracellular loop (ICL3). ICL3 and the C-terminal domain each contain a nucleotide-binding domain (NBD, orange). (B) Mutations in PgABCA2 cDNAs from CRISPR-R2 (3–8 clones from each of five individuals: 3, 5, 9, 10 and 18) relative to the susceptible strain APHIS-S (MG637361). Individuals with two distinct PCR products cloned are indicated as A or B (e.g., 5A and 5B, etc.). Numbers to the right of the decimal point for each individual indicate the clone sequenced. Exon numbers are shown for APHIS-S. Location of sgRNAs 1–5 are shown as colored bars (sgRNA1, magenta; sgRNA2, cyan; sgRNA3, blue; sgRNA4, yellow; sgRNA5, red). Red triangles above bars indicate premature stop codons, which occur in all sequences except 10.1, 10.21, 10.22, and 10.24. Colors within bars show mutations: orange for insertions and deletions (indels), red for deletions, and green for insertions (Adopted from Fabrick et al., 2021)
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