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. 2020 Dec 17;10:221. doi: 10.1186/s13568-020-01162-9

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Construction of thga3 deletion and complemented mutants. a Thga3 gene deleting strategy. b Thga3 gene complementing vector pKH-KO-Gthga3. c PCR and RT-PCR identification of thga3, hyg, and neo genes in Δthga3 and Rthga3. M, 250-bp ladder. PCR templates were genomic DNA and cDNA of wild-type Th33, genomic DNAs of Δthga3 and Rthga3. d Southern hybridization analysis. Probe A. Thga3 ORF; Probe B. Hyg gene; ProbeC. Neo gene. DNAs of wild-type Th33, Δthga3 and Rthga3 were digested by EcoRI/XhoI