Journal: Microbiology Spectrum

Article Title: Versatile allelic replacement and self-excising integrative vectors for plasmid genome mutation and complementation

doi: 10.1128/spectrum.03387-23

Figure Lengend Snippet: Strains and vectors used in this study

Article Snippet: The zeocin resistance marker ( zeoR ) was amplified from pCRBlunt II-TOPO vector (Invitrogen, Waltham, MA USA) using zeo1 and zeo2 primers and inserted into the Not I- Bam HI-restricted pKOV vector (Addgene plasmid #25769, a gift from George Church) in the same orientation as in chloramphenicol resistance gene ( cmR ). zeo1 was designed by adding the Not I restriction site at the 5′end, and similarly, Sac I, Xho I, and Bam HI restriction sites were incorporated to the 5′end of zeo2 for cloning purposes. lacZ alpha amplified from V308 pDNR MCS lacZ alpha-Donor (Addgene plasmid #11691, a gift from Tony Pawson) using lac1 and lac2 primers was inserted at the Xho I site, and the final vector was designated as pKOV-ZeO (Fig. S1).

Techniques: Clone Assay, Plasmid Preparation, Marker, Expressing

Journal: bioRxiv

Article Title: Maternal Wnt11b regulates cortical rotation during Xenopus axis formation: analysis of maternal-effect wnt11b mutants

doi: 10.1101/2022.02.02.478872

Figure Lengend Snippet: (A,C-D,G,I-J,M-N) Phenotypes (A, G) and in situ hybridization of mesendodermal markers (C, myod1 ; D, szl ; I, wnt11b ; J, wnt8a ; M, sox17a ; N, nodal3 . 1 ) in early neurula plate stage (A-D) and mid/early gastrula stages (G-N) in control heterozygotes (+/Z; left-hand panels). (B,E-F,H,K-L,O-P) Phenotypes (B, H) and in situ hybridization of mesendodermal markers (E, myod1 ; F, szl ; K, wnt11b ; L, wnt8a ; O, sox17a ; P, nodal3 . 1 ) in early neurula plate stage (A-D) and mid/early gastrula stages (G-N) in maternal mutant homozygotes (M/Z; right-hand panels). Dorsal, posterior views (A-F); vegetal views (G-P, dorsal towards top). (Q-S) Real-time RT-PCR analysis of myod1 and szl at stage 12/13 (Q), and sia1 (R) and szl at stage 9 and 10.5 (S). Bars in green indicate the samples used for normalization of relative expression; maternal mutants are coloured in cyan. Error bars in (Q) represent standard error of the mean of two biological replicates; representative experiments are shown for (R-S).

Article Snippet: 1 (from R. Harland; Eco RI/T7), eomes and myod1 (from J. Gurdon; Eco RI/T3 and Bam HI/SP6 respectively), sox17a (from A. Zorn; Asp 718/T3), pitx2c/pbluescript (a gift from M. Blum; Not I/T7) and sizzled/pcs2+ (from M. Kirschner, Addgene plasmid 16688, Bam HI/T3), wnt8a/pcs2+ (XE10, from R. Moon; Addgene plasmid 16865; Bam HI/T3).

Techniques: In Situ Hybridization, Control, Mutagenesis, Quantitative RT-PCR, Expressing