1 2 3 4 5 6 7 8 9 DNA DNA DNA DNA
7 A B C D E F
7 A Yeast 56 500
7 A FDA Generally Recognized As Safe GRAS
7 B
Saccharomyces cerevisiae Kluyveromyces lactis Pichia pastoris Schizosaccharomyces pombe Hansenula polymorpha
ssc1 ATP ssc2 rgr1 ose1 ssc11 Y rho -
mnn alg och
Lys HOOC Ubiquitin 76 aa ubiquitin ligase E3 Lys ubiquitin ligase E3 Lys
UBI 1-52 CEP52 UBI 2-52 CEP52 UBI 3-76 CEP76 UBI 4 UBC 1 UBC 2 UBC 3 UBC 4 UBC 5 UBC 6 UBC 7
UBI 4 UBI 4 UBI 4 - UBI 4 UBA 1 UBA1 E1 UBA1 Ubc4 - ubc5
7 C
2µ 2µ 50 100 IRs 600 bp FLP IRs REP STB REP RAF STB REP1 A ori IR IR FLP REP2 psr1 psb1 pkd1 2µ B
pgkl1 pgkl1 DNA αβ γ 50-100 K1 K2 pgkl1 8.9 kb α β γ
ARS YRp ARS 0.8-1.5kb 30-40kb ARS DNA ARS YRp 2µ YEp 200 50%-70%
CEN YCp CEN DNA CEN DNA ARS YCp YCp 1-5 TEL YAC
DNA YIp DNA Yip DNA DNA
7 D
Ca 2+ PEG 1-2% 25-33%
Li + PEG DNA DNA DNA 80
DNA PEG 10 5 / µg DNA
DNA 10-30 YIp DNA 50-80%
LEU TRP HIS LYS URA ADE
aph G418 cat dhfr cup1 suc2 ilv2
7 E
pho4 TS -PHO5 PHO5 PHO4 PHO5 PHO4 pho4 TS 35 pho4 TS -PHO5 35 23
a α a α 25 Sir3-8 TS a1 a MATa MATα hmlα2-102 MATa α α1α α2a a1-a2 α 35 Sir3-8 TS α2 a1 a α2 hmlα2-102 α2 a1 a hmlα2-102 MATa α1 α
GAL1 GAL1 GAL7 GAL7 GAL10 GAL10 UAS UAS GAL4 GAL4 GAL80 GAL80 GAL1 GAL1 GAL7 GAL7 GAL10 GAL10 UAS UAS GAL4 GAL4 GAL80 GAL80 GAL4 GAL1 GAL1 GAL10 GAL4 GAL1 GAL1 GAL10 GAL10 GAL10 Promoter Promoter GAL1 GAL1 GAL7 GAL7 GAL10 GAL10
mrna mrna -3- GAPDH PKG ADH 96% 25
-3- GAPDH PKG ADH
pkd1 40
AOX1 DNA 20
HARS HARS DNA 100
7 F HBV 2003HBV
7 F
DNA 42 nm 3.2 kb HBsAg S HBcAg 22 nm 1000S M L
108 aa 55 aa 226 aa ATG ATG ATG TAA pres1 pres2 S 226 aa S 281 aa M 399 aa L
20 80 HBsAg S ADH1 22 nm HBsAg 1%-2% M L S S
PARS2 Bgl II 5 AOX1 HBsAg pbsag151 11 kb Bgl II 3 AOX1 Bgl II his - PHIS4 DNA his + 5 AOX1 HBsAg PHIS4 3 AOX1
DNA AOX2 HBsAg S 3% 240L 90 22nm HBsAg 900