Table 2 Vectors and strains used in this study

Plasmid

p390-blpR-sgRNA-cmcas9-gfp

Agrobacterium-mediated transformed vector with TrpC promoter, Cmgpd promoter, glufosinate-ammonium resistant selection marker Blp^R, Km^R

–

[26]

pET-CmEgt1-CmEgt2

Carrying CmEgt1 and CmEgt2 at NcoI and XhoI sites, these genes were drove by one T7 promoter and separated by ribosome bind site (RBS), Km^R

pET28a

Lab collection

pET-CmE1b-E2

Carrying CmE1b and CmEgt2 at NheI and XhoI sites, these genes were severally droved by different T7 promoter, Km^R

pET28a

Lab collection

pBAD-EgtD

Carrying EgtD at SacI and HindIII sites, Cm^R

pBAD-Myc-His

Purchase from Thermo fisher Scientific, MA, USA

p390-blpR-CmEgt2

Inserting the CmEgt2 into XbaI site, to build the expression cassette PtrpC-CmEgt2-TtrpC, Blp^R, Km^R

p390-blpR-sgRNA-cmcas9-gfp

[26]

CmE1b_EgtD

Linking CmE1b and synthesized EgtD [30] with a self-cleaving 2 A peptide [36] to build cassette CmE1b-P2A-EgtD

Synthesized

Lab collection

p390-CmEgt2-E1B

Inserting the CmE1b into PstI and BcuI sites, to build the expression cassette Pcmgpd-CmE1b-Tnos, Blp^R, Km^R

p390-blpR-CmEgt2 and

CmE1b-EgtD

This study

p390-CmEgt2-E1b_EgtD

Inserting the CmE1b_EgtD into BcuI site, to build the expression cassette Pcmgpd-CmE1b_EgtD-Tnos, Blp^R, Km^R

p390-blpR-CmEgt2 and CmE1b-EgtD

This study

Strain

BL21-pET-CmEgt1_Egt2

Carrying the vector pET-CmEgt1-CmEgt2, Km^R

E. coli BL21 (DE3)

Lab collection

BL21-pET-CmE1b2

Carrying the vector pET-CmE1b2, Km^R

E. coli BL21 (DE3)

Lab collection

BL21-pBAD-EgtD

Carrying the vector pBAD-EgtD, Cm^R

E. coli BL21 (DE3)

Lab collection

BL21-EgtD1b2

Carrying the vectors pET-CmE1b2 and pBAD-EgtD, Cm^R, Km^R

E. coli BL21 (DE3)

Lab collection

Cordyceps militaris CM15

Wild type strain, with slender and long fruiting body

-

Purchase from Shandong, China

Cordyceps militaris CMdf

Degenerated strain of CM15 which was breeding by continued cultured and showed dwarf and oval shape fruiting body

Cordyceps militaris CM15

Lab collection

15-E1b2

With overexpression boxes of CmE1b and CmEgt2 from p390-CmEgt2-E1B in genome of CM15, Blp^R

Cordyceps militaris CM15

Lab collection

15-E1b2D

With overexpression boxes of CmE1b and CmEgt2 from p390-CmEgt2-E1b_EgtD in genome of CM15, Blp^R

Cordyceps militaris CM15

Lab collection

Df-E1b2

With overexpression boxes of CmE1b and CmEgt2 from p390-CmEgt2-E1B in genome of CMdf, Blp^R

Cordyceps militaris CMdf

Lab collection

Df-E1b2D

With overexpression boxes of CmE1b and CmEgt2 from p390-CmEgt2-E1b_EgtD in genome of CMdf, Blp^R

Cordyceps militaris CMdf

Lab collection