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e￥?aé ? ?é ?ˉ?V?yN￥ù?1
% )% = "
? ? N?
e ￥?aé? ?é ?ˉ?V?yN￥ù?1
í?￥?aé bd8?￥?aéü
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?
?yN31 8?é? |
í?￥?aé bd8?￥?aéü
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d8￥M?Dˉ??T
d8￥ ·3DM?
d8￥ ·3DM?
d8￥ ·3DM?￥?1 ?
^· 7p ￥￥=
;o?Q)o
d8￥ ·3DM?￥?1 ?
^· 7p ￥￥ =
;o ? Q)o
|yHq/
Pd8 ·3i× ?é ?ˉ?o?? b ?μr\é
|yHq/
Pd8 ·3i× ?é ?ˉ?o?? b ?μr\é
d8 ·3M?￥
k4?Hq ?
d8 ·3M?￥
k4?Hq ?
b!4 SDS a?
E=??
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a?
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d8￥ˉ?D×|?
d8￥ˉ?D×|?￥?1 ?
^
d8￥ˉ?D×|?￥?1 ?
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YVQ)o￥??
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d8ˉ??T￥ZE ?
s
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°¤Vrs÷?a?? V|?÷yD
s8 )1
&￥?a
éI
?y ó¤ B iT1B? 7b? O é?Vr b??
??yVr￥?aé?1 μ??a b ?? μ??a2?
Yè
s8% )￥?a?sê?
s÷?aê?
￡
ü
|y
!9? ?￥?a
? Méê ??D
k4+s?
ê? V
[ 8?VB?￥ μ??as0?
dbí
ls÷?a
μ??s÷?a￥Vr
μ??s÷?a￥Vr
[ μ??
TVr "￥?a￥a ??
[ μ??
TVr "￥?a￥a ??
"￥?a×??ú - s0
?l￥ trT÷D
"￥?a×??ú - s0
?l￥ trT÷D
"￥?a^?s ? ?¨
s8?a?
?￥ F8 a ￥8 a?té
"￥?a^?s ? ?¨
s8?a?
?￥ F8 a ￥8 a?té
"￥?aVr
qú D
s8?a ¨B*? ?￥Vríq
"￥?aVr
qú D
s8?a ¨B*? ?￥Vríq
=??
z￥ bW` Ov μ
￡ ·?
=??
z￥ bW` Ov μ
￡ ·?
"￥?a b
L=
3á?á ??1￥?'aaü ?y
"￥?a b
L=
3á?á ??1￥?'aaü ?y
? T?a V[ y
?¤B?ú￥ μ??a
? T?a V[ y
?¤B?ú￥ μ??a
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s8?a￥i μ??aaa ?
"￥?a ·3?z ??
s8?a￥i μ??aaa ?
"￥?a31í
l μ??a31
3?éB?s ?? ?
"￥?a31í
l μ??a31
3?éB?s ?? ?
μ??s÷?a￥Vr
μ??s÷?a￥Vr
μ?? "￥?aVr"d￥y
μ?? "￥?aVr"d￥y
μ??aVré ì￥ye5
μ??aVré ì￥ye5
s8%￥2y ?úrVr O Vrát V[YV T?
s8%￥2y ?úrVr O Vrát V[YV T?
aé?+s?e?B?
aé?+s?e?B?
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1 "￥?as ?í
l7/Hq
1 "￥?as ?í
l7/Hq
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s8?aI
?y￥/31 μ??a4?
?÷y?
s8?aI
?y￥/31 μ??a4?
?
á
?0 b t f ?/i?31??￥
s82y "
?
á
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s82y "
￥
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? μ??as0?
?Fz￥s0
V?¤í
￥
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? μ??as0?
?Fz￥s0
V?¤í
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3y μ??a￥
3y
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??
? ?Bá￥Jr O
??aI
??
? ?Bá￥Jr O
μ??s÷?a￥Vr
μ??s÷?a￥Vr
μ?? "￥?aVr"d￥y
μ?? "￥?aVr"d￥y
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s8?a
¨? μ??ay￥
s8?a
!??tM
?
GST ??
z bW`
!??tM
?
??
z bW`
; ??e?a
TrxA ??
z bW` pTrxFus
; ??e?a
??
z bW`
?è2??a
MBP \és
?
?è2??a
\és
?
á? ? ?$ o )?a A
SAPA
?f T?a pRIT2T
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?f T?a
? ?a
OmpF \és
?
? ?a
\és
?
β-? ?
?
LacZ
?f T?a
? ?
?
?f T?a
W
í?a
Ubi ??
z bW`
W
í?a
??
z bW`
μ??s÷?a￥Vr
μ??s÷?a￥Vr
"￥?a￥í
l
"￥?a￥í
l
μ??a?
s8?a?s￥i V ???Y "￥?a￥ b
W`?
3t? ?T|-? ? |8???á
?fQ?y
N ?!?
3á0¨ "￥?a
H| μ??a?￥
s8?a?
s??" ?
^A? V
￥y? b μ??a￥ê?B?
Z
Eμ
?
?D
E
?3E
μ??a?
s8?a?s￥i V ???Y "￥?a￥ b
W`?
3t? ?T|-? ? |8???á
?fQ?y
N ?!?
3á0¨ "￥?a
H| μ??a?￥
s8?a?
s??" ?
^A? V
￥y? b μ??a￥ê?B?
Z
Eμ
?
?D
E
?3E
μ??s÷?a￥Vr
μ??s÷?a￥Vr
"￥?a￥í
l
"￥?a￥í
l
¨??aê?B??D
￥KD
k41? e
CNBr
?D t y?￥J
;?
??§ y ￥
;
?Q?
3??e? =
a??×?
￡￥T¨/t o
??
? t?3 ?
?
3t
￥J
;?
??? 1ú
′?
??7/3t
N
￥?Bê?
?? ??M b ?BZE￥a?
^í
l
qú
V
r? 85%[
B? <7 O
?á
3￥ "￥?a￥ N
?cJ
;?
?D??
3t%?￥?
? Vrát?1¤í b ?7 ?T
s÷?as0 =?cμJ
;?
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￥KD
k41 ? e
?D t y?￥ J
;?
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;
? Q?
3??e? =
a??×?
￡￥T¨/t o
??
? t?3 ?
?
3t
￥J
;?
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′?
? ?7/3t
￥?Bê?
?? ??M b ?BZE￥a?
^í
l
qú
V
r? [
B? <7 O
?á
3￥ "￥?a￥
?cJ
;?
?D??
3t%?￥?
? Vrát?1¤í b ?7 ?T
s÷?as0 =?cμJ
;?
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?D
E
?D
E
μ??s÷?a￥Vr
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l
"￥?a￥í
l
?3E ??ê?
?3E ??ê?
?a = M
?
?a = M
?
Méê?
Méê?
è )?a
? Arg-C
è )?a
?
?$ o )?a
? Glu-C
?$ o )?a
?
L?1 )?a
? Lys-C
L?1 )?a
?
ìO?a
? Arg-C
ìO?a
?
Lys-C
?a
?
?3E￥+?
^
r
q÷ú
?a
?
?3E￥+?
^
r
q÷ú
]
H
???a
? ( μM?￥
ê? %?
]
H
???a
? ( μM?￥
ê? %?
[yN Vê4￥B?
ê?S??
[ yN Vê4￥B?
ê?S??
< b+?
ê?B?K <￥
???a
< b+?
ê?B?K <￥
???a
?sY t y?￥ú?
? a!?
? a ??
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??￥
? ?) M 7 ??o???c
??￥
/3
t
D? e?D
EM
? b
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t
D? e?D
EM
? b
¨
??a
?
3 μ??a￥ -4Hq
^?
¨
??a
?
3 μ??a￥ -4Hq
^?
÷?as0 =?? ?cμú?
? a!?
?
÷?as0 =?? ?cμú?
? a!?
?
??
?? ?T?÷yVrát1ls0 t?BK?Hqi? N Y?
??
?? ?T?÷yVrát1ls0 t?BK?Hqi? N Y?
vs0
￥s÷?a ?
a
? ?? ?
??￥C ?
q
^M?ú￥
vs0
￥s÷?a ?
a
? ?? ?
??￥C ?
q
^M?ú￥
Arg CN N C
s8?a
s8?a
"￥?a
"￥?a
μ??s÷?a￥Vr
μ??s÷?a￥Vr
"￥?a￥í
l
"￥?a￥í
l
?3E ?ê?
?3E ?ê?
1
X?? M?B?
?? ￥?a
?
?{ ?￥?¨ K?
V
s8?aI
??
D?c
S
á
?0￥?÷y-WF
B
I
?*t?
Ile-Glu-Gly-Arg￥ |y¤h ?
?*t1 μ
?a
??￥ `óy0 9 B￥
MY?T¨?
ê? Arg￥
C
bB?a￥ μ??a¨ Xa) ?' V¤?c
?*t?
￥ "￥?a b?? Xa￥
MYT¨?
?
1??
??F?v
??aé?C??*t?
￥ à
q
yN??ZE V<W
¨?V μ??a?í
lò??]vl￥ "￥?aát
1
X?? M?B?
?? ￥?a
?
?{ ?￥?¨ K?
V
s8?aI
??
D?c
S
á
?0￥?÷y-WF
B
I
? *t?
￥ |y¤h ?
?*t1 μ
?a
??￥ `óy0 9 B ￥
MY?T¨?
ê?
bB?a￥ μ??a¨ ) ?' V¤?c
?*t?
￥ "￥?a b?? ￥
MYT¨?
?
1??
??F?v
??aé?C??*t?
￥ à
q
yN??ZE V<W
¨?V μ??a?í
lò??]vl￥ "￥?aát
μ??s÷?a￥Vr
μ??s÷?a￥Vr
"￥?a￥í
l
"￥?a￥í
l
?3E ?ê?
?3E ?ê?
?0
?0
s8y
s8y
¤h
¤h
"￥y
"￥y
Met Arg Stop
Lys
Glu
Ile-Glu-gly-Arg
Arg
Lys
Glu
Ile-Glu-gly-Arg
Vr
Vr
T?a
T?a
?3í
l
?3í
l
μ??s÷?a￥Vr
μ??s÷?a￥Vr
"￥?a￥í
l
"￥?a￥í
l
?3E ?ê?
?3E ?ê?
Pinpoint Xa
tac
3t
í2?tI
??
3t
í2?tI
??
Xay0
MYê?I
??
y0
MYê?I
??
SP6
T7
Ap
r
r
ori
MCS
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* M F ( M V ( M Z " S H ( M V
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NruI HindIII PvuII BamHI KpnI EcoRV BglII SmaI NotI
Xay0 Méê?
y0 Méê?
*
?s÷?a￥Vr
*
?s÷?a￥Vr
V ?
?
??÷y￥Vr
￡ üD
s8%? V
cy￥ J
?
'y4
??M1 b ? 7×Fé ì"
cμ?÷y?
?z{
?￥ V
cy ?T1 ?êS:￥ F
3
í F?y? b
?
"×Fé ì J
?￥?9F
s 8% =￥v?s ?
?e
$¨
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?μ￥×Fé ìI
??a 7%￥?è
3é}? ?y ?
?
6
s??YyNYV9Fé ì J
?4ú?÷yVrát￥á
aa? ?¤
?i￥rT b
6B?YV9F?÷y4
74ú "S?aá
￥μrZE
^
y*
1 ó?s÷?aVr8 '| J ￥?÷y X
H B
?? J é ì
[ |}? J ? ÷y ú J 8
Vr￥|
{
V ?
?
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￡ üD
s8%? V
cy￥ J
?
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cμ?÷y?
?z{
?￥ V
cy ?T1 ?êS:￥ F
3
í F?y? b
?
8% =￥v?s ?
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??a 7%￥?è
3é}? ?y ?
?
6
s??YyNYV9Fé ì J
?4ú?÷yVrát￥á
aa? ?¤
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6B?YV9F?÷y4
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￥μrZE
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y *
1 ó?s÷?aVr8 '| J ￥?÷y X
H B
?? J é ì
[ |}? J ? ÷y ú J 8
Vr￥|
{
*
?s÷?a￥Vr
*
?s÷?a￥Vr
[*
?
TVr "￥?a￥a ??
[*
?
TVr "￥?a￥a ??
"￥?aúrVr
"￥?aúrVr
?4úé ì J
?￥ -4/9F "￥y￥ J
? V[
?4úé ì J
?￥ -4/9F "￥y￥ J
? V[
×?Vrls0 t
×?Vrls0 t
B??
? ?Vr
B??
? ?Vr
"￥átí
l ? 4
"￥átí
l ? 4
t?? ?Cμr￥ bW2 % )%?￥?
? ù? b
t?? ?Cμr￥ bW2 % )%?￥?
? ù? b
1 ó t μD?aéM
?￥é# bW2y7 F?a
?
1 ó t μD?aéM
?￥é# bW2y7 F?a
?
?3￥ ? ?v?4ú
?3￥ ? ?v?4ú
*
t31
3?éB?s ?? ?K?s0?
3a
*
t31
3?éB?s ?? ?K?s0?
3a
￥ ts0 ?^C?
? (B?
￥ ts0 ?^C?
? (B?
*
?s÷?a￥Vr
*
?s÷?a￥Vr
*
? "￥?aVr"d￥y
*
? "￥?aVr"d￥y
P SD gene gene gene gene T
1
T
2
1
H
2
N COOH
Met Met Met Met
mRNA
CNBr
'^
{
'^
{
*
?s÷?a￥Vr
*
?s÷?a￥Vr
*
? "￥?aVr"d￥y
*
? "￥?aVr"d￥y
y è
y è
P SD T
1
T
2
1
EcoRI BamHI
AATTCAGATCT
GTCTAGA
G
CCTAG
EcoRI Bgl II BamHI
EcoRI BamHI
Bgl II
GATCT
A
G
CCTAG
EcoRI BamHI
Bgl II
EcoRI BamHI
Bgl II
EcoRI + BamHI
Bgl II + BamHI
BamHI
???s÷?a￥Vr
???s÷?a￥Vr
[???
TVr "￥?a￥a ??
[???
TVr "￥?a￥a ??
"￥y×?Vr
"￥y×?Vr
???￥ "￥y
?
s8% ? ?8 DNA￥ˉ?7ˉ? v
???￥ "￥y
?
s8% ? ?8
? f ?/M?×?y? )
àμê4a ?i / V[ ??
? f ?/M?×?y? )
àμê4a ?i / V[ ??
"￥yVr
q?
"￥yVr
q?
? J ??￥ "￥yVr
q
s?K?N
H VYV <?Vr
? J ??￥ "￥yVr
q
s?K?N
H VYV <?Vr
íq7F[?ê
íq7F[?ê
?!7?é
> "￥yVr??[?
t?L.?1 "
?!7?é
> "￥yVr??[?
t?L.?1 "
￥￥yy?? è+Yμil
￥￥yy?? è+Yμil
???s÷?a￥Vr
???s÷?a￥Vr
DNA8 =×F￥'e ?
8 =×F￥'e ?
3t8%-
^e?% )% =<Wi " DNA
￥L.×F? V ?￥
3tD?r
^\é
3t? ?￥é
? b% =￥L.×F Vs1
v ?
3t8%-
^e?% )% =<Wi "
￥L.×F? V ?￥
3tD?r
^\é
3t? ?￥é
? b% =￥ L.×F Vs1
v ?
êy0G ??
êy0G ??
]÷?
G ??
]÷?
G ??
???s÷?a￥Vr
???s÷?a￥Vr
DNA8 =×F￥'e ?
8 =×F￥'e ?
êy0
^·
3t% =? ?i ￥B ?íˉ? ? ?￥ DNA V
M?y0?]
3t? ??μ2?]￥êy0 è ?
êy0
^·
3t% =? ?i ￥B ?íˉ? ? ?￥
M?y0?]
3t? ??μ2?]￥êy0 è ?
êy0G ??￥8 =×F êy0EB
êy0G ??￥8 =×F êy0EB
` )8 G ?
G-Fragment
` )8 ?
e?% ) ? ?
¨?
IS
e?% ) ? ?
¨?
??% ) V0
Tn aTy
??% )
ú?±t VM?y0
Ac aDs
ú?±t M?y0
ú??t Ky
mobil gene
ú??t
???s÷?a￥Vr
???s÷?a￥Vr
DNA8 =×F￥'e ?
8 =×F￥'e ?
êy0G ??￥8 =×F êy0￥2
êy0G ??￥8 =×F êy0￥2
IS
1 kb
Ty
5 kb
Tn
2 - 20 kb
ACCATGGTT TTGGTACCA
F0?y
F0?y
ê
?y
ê
?y
MYê?
MYê?
E3y
E3y
IR IR
IR IR
IS IS
???s÷?a￥Vr
???s÷?a￥Vr
DNA8 =×F￥'e ?
8 =×F￥'e ?
êy0G ??￥8 =×F ???
T
êy0G ??￥8 =×F ???
T
???s÷?a￥Vr
???s÷?a￥Vr
DNA8 =×F￥'e ?
8 =×F￥'e ?
? e?% )%? ? ?8 DNA y
￥
?]÷ u-W V
?
3
?ì￥]÷×F ?
qD% ) ? ? a
?]÷ u-W￥ ?
]÷ u￥é a]÷?
á MM1 b B?1
a ? ù aé
é a]÷? ú×F￥ ?
q9ü úQ-g ? b
]÷×Fμ????D
??
T -?o31B?
ê?
7a?
#
?
ê?
? e?% )%? ? ?8 y
￥
?]÷ u-W V
?
3
?ì￥]÷×F ?
qD% )
?]÷ u-W￥ ?
]÷ u￥é a]÷?
á MM1 b B?1
a ? ù aé
é a]÷? ú×F￥ ?
q9ü úQ-g ? b
]÷×Fμ ?? ? ?D
??
T -?o31B?
ê?
7a?
#
?
ê?
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G ??￥8 =×F '?
T
]÷?
G ??￥8 =×F '?
T
???s÷?a￥Vr
???s÷?a￥Vr
DNA8 =×F￥'e ?
8 =×F￥'e ?
]÷?
G ??￥8 =×F ]÷??
]÷?
G ??￥8 =×F ]÷??
ori
"￥y
"￥y
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]÷ u×
S:y
S:y
??ê?
??ê?
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? ?8
???s÷?a￥Vr
???s÷?a￥Vr
DNA8 =×F￥'e ?
8 =×F￥'e ?
]÷?
G ??￥8 =×F ]÷?D
]÷?
G ??￥8 =×F ]÷?D
ori
"￥y
"￥y
]÷ u×
]÷ u×
S:y
S:y
?D u×
?D u×
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? ?8
ori
S:y
S:y
?a
? F? ?J?Vr"d￥y
?a
? F? ?J?Vr"d￥y
í
?
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3t?
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ra??
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￥
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v
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s8%?a
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LV
ü
×F "￥?a
s8% =￥?
? ù V[YV?a?
￥ |
y
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s8%￥?/F[??? e? b
í
?
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| 课件名称： | 华东理工大学：基因工程 |
| 课件分类： | 生物 |
| 课件类型： | 电子教案 |
| 文件大小： | 3.05MB |
| 下载次数： | 2 |
| 评论次数： | 2 |
| 用户评分： | 7.5 |
