Research Article
Development of the Multiple Gene Knockout System with One-Step PCR in Thermoacidophilic Crenarchaeon Sulfolobus acidocaldarius
Table 1
Strains and DNA used in this study.
| | Strain or plasmids | Relevant characteristic(s) | Source or reference |
| | Strains | | | | S. acidocaldarius | | | | SK-1 | MR31 [32] with ΔsuaI | [8] | | DP-1 Int | SK-1 with Δphr::3 region of phr-pyrE-lacS | This study | | DP-1 | SK-1 with Δphr | This study | | SK-5 Int | SK-1 with ΔargD::5 region of argD-pyrE-lacS | This study | | SK-5 | SK-1 with ΔargD | This study | | Plasmids | | | | pSAV2 | Sulfolobus-E. coli shuttle vector, based on pBluescript II KS (−) and pRN1, with the SsopyrEF marker | [8] | | pSAV2-argD | SsopyrEF marker in pSAV2 replaced by SsoargD marker | This study | | pSuaIPOP | pBluescript II KS (−) carrying the 800 bp of 5 and 3 regions of suaI, pyrE, and 800 bp of 3 region of suaI | [8] | | placSpyrE | pSuaIPOP derivative carrying 800 bp of 5 and 3 homologous regions of suaI locus at both ends of pyrE-lacS dual marker | This study | | PCR product | | | | pyrElacS800 | Linear DNA carrying 800 bp of 5 and 3 homologous regions of suaI locus at both ends of pyrE-lacS dual marker | This study |
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