Strains

2.1 Strains

Table 2 contains the strains used and generated for this study, ordered by organism.

Table 2. Strains generated and used for this study

Name Genotype Source

Escherichia coli

Mach1 ΔrecA139 endA1 tonA Φ80(lacZ)ΔM15 ΔlacX74 hsdR(rK- mK+), recipient strain for vector amplification

Invitrogen, Germany

Saccharomyces cerevisiae

PJ69-4A MATa trp1-901 leu2-3_112 ura3-52 his3_200 ga14Δ ga18OΔ LYS2::GALl-HIS3 GAL2-ADE2 met2::GAL7- lacZ

James et al.,1996

Y187 MATα; ura3-52; his3-200; ade2-101; trp1-901; leu2-3, 112; gal4Δ; metΔ; gal80Δ; MEL1; URA3::GAL1UAS- GAL1_TATA-lacZ

Clontech, Mountain View, USA

AH109 MATa; trp1-901; leu2-3, 112; ura3-52; his3-200; ade2- 101; gal4Δ; gal80Δ; LYS2::GAL1UAS-GAL1_TATA-HIS3;

MATα; ura3-52; his3-200; ade2-101; trp1-901; leu2-3,112; gal4Δ; metΔ; gal80Δ;MEL1;URA3::GAL1UAS -GAL1_TATA-lacZ; GAL4-binding domain N-terminally fused SmGPI1 from Smgpi1 cDNA encoding aa 1-253 under control of adh promoter

this study

Y187 + pBD-SmGPI1 aa 28-227

MATα; ura3-52; his3-200; ade2-101; trp1-901; leu2-3,112; gal4Δ; metΔ; gal80Δ;MEL1;URA3::GAL1UAS -GAL1_TATA-lacZ; GAL4-binding domain N-terminally fused SmGPI1 from Smgpi1 cDNA encoding aa 28-227 under control of adh promoter

this study

Y187 + pBD-SmGPI1 aa 1-100

MATα; ura3-52; his3-200; ade2-101; trp1-901; leu2-3,112; gal4Δ; metΔ; gal80Δ;MEL1;URA3::GAL1UAS -GAL1_TATA-lacZ; GAL4-binding domain N-terminally

this study

36

fused SmGPI1 from Smgpi1 cDNA encoding aa 1-100 under control of adh promoter

Y187 + pBD-SmGPI1 aa 101-253

MATα; ura3-52; his3-200; ade2-101; trp1-901; leu2-3,112; gal4Δ; metΔ; gal80Δ;MEL1;URA3::GAL1UAS -GAL1_TATA-lacZ; GAL4-binding domain N-terminally fused SmGPI1 from Smgpi1 cDNA encoding aa 101-253 under control of adh promoter

this study

AH109 + pAD-SmMOB3

MATa; trp1-901; leu2-3, 112; ura3-52; his3-200; ade2 - 101; gal4Δ; gal80Δ; LYS2::GAL1UAS-GAL1_TATA-HIS3;

GAL2_UAS-GAL2_TATA-ADE2; URA3::MEL1_UAS -MEL1_TATA- lacZ; MEL1; GAL4-activation domain N-terminally fused to SmMOB3 from Smmob3 cDNA coding for SmMOB3 full-length under control of adh promoter

this study

AH109 + pAD-SmMOB3 aa 1-144

MATa; trp1-901; leu2-3, 112; ura3-52; his3-200; ade2- 101; gal4Δ; gal80Δ; LYS2::GAL1UAS-GAL1_TATA-HIS3;

GAL2_UAS-GAL2_TATA-ADE2; URA3::MEL1_UAS-MEL1_TATA- lacZ; MEL1; GAL4-activation domain N-terminally fused to SmMOB3 from Smmob3 cDNA coding for SmMOB3 aa 1-144 under control of adh promoter

this study

AH109 + pAD11 aa 282-845

MATa; trp1-901; leu2-3, 112; ura3-52; his3-200; ade2- 101; gal4Δ; gal80Δ; LYS2::GAL1UAS-GAL1_TATA-HIS3;

GAL2_UAS-GAL2_TATA-ADE2; URA3::MEL1_UAS-MEL1_TATA- lacZ; MEL1; GAL4-activation domain N-terminally fused to PRO11 from pro11 cDNA encoding aa 282-845 under control of adh promoter

this study

AH109 + pAD11FL MATa; trp1-901; leu2-3, 112; ura3-52; his3-200; ade2- 101; gal4Δ; gal80Δ; LYS2::GAL1UAS-GAL1TATA-HIS3;

GAL2_UAS-GAL2_TATA-ADE2; URA3::MEL1_UAS-MEL1_TATA- lacZ; MEL1; GAL4-activation domain N-terminally fused to PRO11 from pro11 full-length cDNA under control of adh promoter

this study

Y187 + pBD-SmKIN3 MATα; trp1-901; leu2-3, 112; ura3-52; his3-200; ade2- 101; gal4Δ; gal80Δ; LYS2::GAL1UAS-GAL1_TATA-HIS3;

GAL2_UAS-GAL2_TATA-ADE2; URA3::MEL1_UAS-MEL1_TATA- lacZ; MEL1; GAL4-binding domain N-terminally fused to SmKIN3 from Smkin3 full-length cDNA under control of adh promoter

this study

37

Y187 + pBD-SmKIN24 MATα; trp1-901; leu2-3, 112; ura3-52; his3-200; ade2- 101; gal4Δ; gal80Δ; LYS2::GAL1UAS-GAL1_TATA-HIS3;

GAL2UAS-GAL2TATA-ADE2; URA3::MEL1UAS-MEL1TATA- lacZ; MEL1; GAL4 activation domain N-terminally fused toSmKIN24 from Smkin24 full-length cDNA under control of adh promoter

this study

Sordaria macrospora

S48977 wild type U. Kück, Bochum^a

S66001 Δku70::nat^R; fertile Pöggeler & Kück, 2006

S23442 mutation in fus1-1 gene; brownish ascospores Nowrousian et. al., 2012 S67813 mutation in gene r; pink ascospores U. Kück, Bochum^a

ΔSmkin3 ΔSmkin3::hyg^R, ssi, fertile this study

ΔSmkin3/R2 ΔSmkin3::hyg^R, pink ascospores, ssi, fertile this study

ΔSmKIN3::pRS-SmKIN3+

ΔSmkin3::hyg^R, pRS-SmKIN3+^ect, ssi, fertile this study

ΔSmkin3::pDS-SmKIN3ngfp

ΔSmkin3::hyg^R, pDS-SmKIN3ngfp^ect, ssi, fertile this study

ΔSmkin24 ΔSmkin24::hyg^R, ssi, sterile this study

ΔSmkin24/R2 ΔSmkin24::hyg^R, pink ascospores, ssi, sterile this study

ΔSmkin24::pRS-SmKIN24+

ΔSmkin24::hyg^R, pRS-SmKIN24+^ect, ssi, fertile this study

ΔSmkin24::pDS-SmKIN24ngfp

ΔSmkin24::hyg^R, pDS-SmKIN24ngfp^ect, ssi, fertile this study

ΔSmkin3/ΔSmkin24 ΔSmkin3::hyg^R, ΔSmkin24::hyg^R, ssi, sterile this study S48977::pHA11 nat^R, pHA11^ect, ssi, fertile this study

S67813::pFLAG-SmKIN3

pink ascospores, hyg^R, pFLAG-SmKIN3^ect, ssi, fertile this study

S48977::pFLAG-SmKIN3, S48977::pHA11

pFLAG-SmKIN3^ect, pHA11^ect, nat^R, hyg^R, ssi, fertile this study

ΔSmgpi1 ΔSmgpi1::hyg^R, ssi, fertile Bernhards 2010

ΔSmgpi1/r2 ΔSmgpi1::hyg^R, pink ascospores, ssi, fertile this study ΔSmmob3 ΔSmmob3::hyg^R, ssi, sterile Bernhards & Pöggeler,

2011 ΔSmmob3/r2 ΔSmmob3::hyg^R, pink ascospores, ssi, sterile this study ΔSmgpi1/ΔSmmob3 ΔSmgpi1::hyg^R, ΔSmmob3::hyg^R, ssi, fertile this study

38

ΔSmgpi1/ΔSmmob3/r2 ΔSmgpi1::hyg^R, ΔSmmob3::hyg^R, pink ascospores, ssi, fertile

this study

ΔSm3978 ΔSm3978::hyg^R, ssi, fertile this study

ΔSmmob3/ΔSm3978 ΔSmmob3::hyg^R, ΔSm3978::hyg^R, ssi, sterile this study Δpro11 Δpro11::hyg^R, ssi, sterile Bernhards & Pöggeler,

2011 ΔSmgpi1/Δpro11 ΔSmgpi1::hyg^R, Δpro11::hyg^R, ssi, sterile this study

Δpro22 Δpro22::hyg^R, ssi, sterile Bloemendal et al., 2012

ΔSmgpi1/Δpro22 ΔSmgpi1::hyg^R, Δpro22::hyg^R, ssi, sterile this study

Δpro45 Δpro45::hyg^R, ssi, sterile U. Kück, Bochum^a

ΔSmgpi1::hyg^R, pRS-SmGPI1-egfp aa 1-27-KDEL^ect, nat^R, ssi, fertile

S48977::p1783 S48977, p1783^ect, hyg^R, ssi, fertile Voigt & Pöggeler, 2013 S48977::pMOB3-FLAG

S48977::pRS-SmGPI1-egfp aa 1-492

S48977, pMOB3-FLAG^ect, pRS-SmGPI1-egfp aa 1-492^ect, hyg^R, nat^R, ssi, fertile

this study

S48977::pMOB3-FLAG S48977, pMOB3-FLAG^ect, hyg^R, ssi this study

S48977::pRS-SmGPI1-egfp aa 1-492

S48977, pRS-SmGPI1-egfp aa 1-492^ect, nat^R, ssi, fertile this study S48977::p1783-1

S48977 ::pFLAG Mob3

S48977, p1783-1^ect, pFLAG Mob3, hyg^R, ssi this study S48977::pFLAGN1

S48977::pRS-SmGPI1-S48977, pFLAGN1, hyg^R, pRS-SmGPI1-egfp aa 1-492^ect, this study

39

egfp aa 1-492 nat^R, ssi, fertile

ΔSmgpi1::hyg^R, ΔSmmob3::hyg^R, pRS-SmGPI1 aa 28-253^ect, nat^R, ssi, sterile Δpro11/ΔSmkin24 Δpro11::hyg^R, ΔSmkin24::hyg^R, ssi, sterile this study ect = ectopically integrated, FL = full-length, nat = nourseothricin-cassette, hph = hygromycin-cassette, pt = primary transformant, ssi = single spore isolate, hyg^R = hygromycin resistant, nat^R=nourseothricin resistant, amp^R = ampicillin resistant, kan^R= kanamycine resistant ^a Institute of Biochemistry, University of Stuttgart, Germany.

Im Dokument The STRIPAK complex and its role in fruiting-body development of the filamentous fungus Sordaria macrospora (Seite 48-52)