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Table 2

Analysis of Tim23p Deletion Constructs in tim23::URA3 Yeast Cells

YEPDYEPgly/eth
24°C30°C34°C37°C24°C30°C34°C37°C
Vector − − − − − − − − 
TIM23 +++ +++ +++ +++ +++ +++ +++ +++ 
23Δ2-24 − − − − − − − − 
23Δ2-50 +++ +++ +++ +++ ++ ++ − 
23Δ51-94 − − − − − − 
23Δ2-24,75-94 − − − − − − 
23Δ2-94 − − − − − − − − 
YEPDYEPgly/eth
24°C30°C34°C37°C24°C30°C34°C37°C
Vector − − − − − − − − 
TIM23 +++ +++ +++ +++ +++ +++ +++ +++ 
23Δ2-24 − − − − − − − − 
23Δ2-50 +++ +++ +++ +++ ++ ++ − 
23Δ51-94 − − − − − − 
23Δ2-24,75-94 − − − − − − 
23Δ2-94 − − − − − − − − 

Plasmids expressing wild-type Tim23p (pJE50) or the deletion constructs Tim23Δ2-24 (pAD109), Tim23Δ2-50 (pAD105), Tim23Δ51-94 (pAD106), Tim23Δ2-24,75-94 (pAD111), or Tim23Δ2-94 (pKR15) were transformed into tim23::URA3 trp1 leu2 cyh2 strain KRR146, which contains the TIM23-TRP1-CYH2 plasmid pKR1 (Ryan et al. 1998). Leu+ transformants were patched onto SD medium lacking leucine, replica-plated onto YEPD or YEPgly/eth plates containing 10 mg/l cycloheximide to detect the loss of pKR1, and allowed to grow at 24, 30, 34, or 37°C for 3 d. Wild-type growth (+++), intermediate growth (++ or +), or no growth (−) is indicated.

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