Nghiên cứu biểu hiện gen Trehalase tái tổ hợp ở E.coli và xác định các tính chất của enzyme

Khóa luận tốt nghiệp Viện Đại Học Mở
MỞ ĐẦU
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 8C$<=(; 2@2.
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/O 5.2P/.AN
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;:!:!:R2:<=?IC;:! :!4$
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,&7&@2+" 604 (
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
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Khóa luận tốt nghiệp Viện Đại Học Mở
F5#7#68?IC:R2:;:!:*U%S
* 2XH*&I3(
;+!%< 6-$68K
“Nghiên cứu biểu hiện gen Trehalase tái tổ hợp ở E.coli và xác định các
tính chất của enzyme”
L=6,$K
Y(Z:;:!:[1E.coli(
\(;#&:R2:(
](BH>^,:R2:
_(`<=:R2:[;:! :(
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
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Khóa luận tốt nghiệp Viện Đại Học Mở
PHẦN 1:TỔNG QUAN TÀI LIỆU
I.1 Công nghệ DNA tái tổ hợp
ab7<$!7 $*b70/? 
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*b7:#C$4!c,(
A2Yde]f<!:#):<:!K*b7<$!*b7<g&
*?A/&[H":V4*:,23DV >
!2AA/?*?A?,234 #4:#
A4!,DV(ab7:/PVA2Ydhh/
i2 *b76:j2$U#/&[: >
:2"+ 2"(ab7:/ i2234*b
70/?!*b7Ek[!7:O:<
:# :(
Ek[!Ek +$i#7!<
$*(lEkO4+ Ek,
* !  >! *Ek[(

6P,Ek[/ i2234/"
(;"&: 2:2=6#Ek?m#: n&#
P+ .23! oR2:&(; $.
m>/4#"#7/7 nEk: 2:&/jEk
0/QmEkn,2fk=(Ek[+*
fk.I-,:R2: Om:#:::
p<::<:Ekp !2::n(Ek#: !23!2< >
:m&#72<X:!#*@n!2<;,#*@
Agrobacterrium tumefaciensm&#72<X:!&/ #7n
!Fmretrovirus >andovirus*/&#72<X
:!&/ 3#7n(Z"8!#7Ek[# 0
#72<X#/"&!C#72?IC
m&C#&n?@22=6(
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
3
Khóa luận tốt nghiệp Viện Đại Học Mở
 .PQ23! :R2:&m.!:R2:V
<^n Ek2=6# Ek#: mUP232
,2#XI P*Ong65!6*&< 0
/Q1V<^i!:PV*(a&D? q
5[,#: #"23 >2342?Ek,
: 2:2=6m/6?@2*2 24#: 
*O#X#: [#"#2?Ek*[
 >[n(;C??@2 q$3$
#"#72 <X#72C 2
.<<G&4!mC*4#"#*@
C&4#"&/ 3#7#7n(; $*U
&/ #72#: 4(r0< !2</H:R2:&
P #X23#H^IH 23:$*233^
7/&*!2<[2C*?A ^1

#72ms2C*?A[Z:p! <:n6U
#724#*/^5
C <X(Z"&: !<X^,:2=6
 ><X:2=6m.!K02
2'<Hotu)kv::2/! ! :/! ! >#"
<XEk >fkw :/! ! :/! !n >
lJf( C <X#7292/"
26- <XV#72?IC?
@22=6(
;C,Ek[!7$*b76
23:?/&C-,:/3: :2" /3:(
JEk[/ i2/"0/?(
p ;&Ekfkm!7:n
p ; #: [m/ i2@/H#: <Xn
p Z&#: [# &/ ,#<X(
p )!: <c 3#/,:
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
4
Khóa luận tốt nghiệp Viện Đại Học Mở
 J &*b7<$/"&#/7(
$?@2^?IC+##7[#
#7/&[:m:!!2 <x:< 2wyLzn4T
470<V/N2:4/ >*
449< >C!?@2 * 8
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<'?ICm<'C6!8C&/ n#<'
<= A4 >3#7[??@2
^4/72^ !2*
6.#!#7 2 >3*$H
../V&3DN;6*b7<

$T>! .# 5?0V#":
<2"!6D"8*9:,.#w2.(J
82^#,yLz@24#" .T
,7*?A!$:*4m,l!2<#: n
 7 #*@6,0m#^<= .3n
.#3#7(JA2! i$*4<9#
4</<g&!2<=<#& 1
9<5#7*4#*/{!230
2"4#"(;00#7yLz!C
#76{!C #<g&<,$! 
#77D?62&2.N
I.2 Đại cương về vi khuẩn E.coli
;: ' FS  l" mYdh]n|  ' tETmYdh}n 
*@ Escherchia Coli   6  *     ! Bacterium Coli Comunis /5
.E.coli-1V,3m3#V4,3 n
^*1<<^"3 ,3#7m'FS
l"Ydhen(;U!E.coli1*&!~e(]•~~(]•2
!]d(]•E.coli IC#413#7U#.*#
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
5
Khóa luận tốt nghiệp Viện Đại Học Mở
i&* #7&(t A#AH?$E.coli0! A
9(;: a:x2,E.coli.>#$&
): :6(; 4234: :#XD
 #/ .#3#7(
I.2.1 Đặc điểm hình thái
E.coli!! *@57PVX!<3
*5/ 2/P2y22.g21
V1*^"#*@\p]LI€(_p€(}2(; 0
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I.2.2 Đặc tính nuôi cấy
E.coli!#*@&*^*H*^Q<'C
1#^C1]h

Jr^!h(\ph(_(;6
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L."HKE.coliC2.=$
!P>I42 62>52292
<^# 42Q2Q4(
L.H.KC1]h

J \_5*@
!X""!4* 42 .*^*@!
* ?\p]22(
L.o< K5*@!*2(
L"))K62.\_C5*@
!2i >29:(
L.oLZKE.coli*@!2^2:(
L.L *:KE.coli5*@!29(
L.Z!!wy::wK*@!,E.coli<)2
#(
I.2.3.Đặc tính sinh hóa
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Khóa luận tốt nghiệp Viện Đại Học Mở
J,E.coli$!62:02! .y! :
y! :t :L !N
E.coli!62:*0! .) :f2 :
B!IF!y!I: (
J?8*K

l?8< !mƒn?8Flm‚n?8Lfmƒn?8r\)
mƒn 6„::mpny:!mpn(
I.2.4 Sức đề kháng của E.coli
E.coli/H6<13~~

J 23.}€

J ]€-
mv!! #L!:Ydd~nm'#Sl"Ydh]n(JCQK
kI:J! ,M 2 !6<E.coli ~-(
I.3. Tình hình sản xuất và ứng dụng enzyme trên thế giới và Việt Nam
I.3.1 Tình hình sản xuất và ứng dụng enzyme trên thế giới
oR2:#X#QD*UI- ?
8I? 442**944
-#gI- ?8 &/ m n(Z1#7
:R2:(0/?#$:R2:!0
1688<=:R2: '(
oR2:.<=: K*:R2:
*96!2 $*7!  3,234:R2:
… 6!-CQ 6
!: "2 24(r >:R2:*96!1<
&@2*V&(oR2: 3+/C*†
0 D 4  ( F^<= :R2:  0 2     ? IC +
Acitinoplanes"Zymomonas(;+#Hmn&3,P(
E 7<=i6!:R2::R2:<Q
:R2:&/&! 6!* ><=# 
2=^*(rA2:R2:02?IC 
023+C224#C22:DY‡]+#*@X!e•+
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Khóa luận tốt nghiệp Viện Đại Học Mở
3#7_•+#7(
F#7!i6!4C,:R2:#5
-43162.<<G*P
$*X9&/HP$3 >$*A
C Nr0,3$*D5[
:R2: 2!:R2: &/ ##7<'<0&
/ #7#3#7(
rA2A2C&@2:R2:?IC=#=
 #(A2Yde€6&"?IC~]€C
 ::+#*@]~€C! 2!:]\€C α wk2!:h€C
!  2::C?HY~€„)E(
r6H.* ?Y\?IC:R2:^#"
6}€C!"(r?IC:R2:!"C!
 #  <*mWLn#yZ <:&2}€•!:R2:6
H.(;&!LbY~• \‡]<=3H,&<Q
?ICi#.H? (A2Ydd]6H.Lb6=~€
„)E4#":\h„)E4#"! k2!:\]„)E
4#"!  2::Na ?~€! :R2:*/6
H.Lb(7&2Y\pY~•H.6=#"?@2
<(;ˆ4#!H.?IC,&=#=
 "(
W&.  /&# ! * ?]~€€! :R2:
* ?#A2! ?IC6D2,&!
:R2:k2!:l ::(t:N
p k2! <:K?IC#"?!~C‡A2<=tp
<2*9qEtI2m\~€CtI26A2n
p k2! ! <:K?!YC‡A2?IC]€€€C! :+
/3‡A2(
p y!  2::K    <=    ?  IC  I ! :  w  x :  _\•

Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
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Khóa luận tốt nghiệp Viện Đại Học Mở
m\(Y~€(€€€C‡A2n#I ! :px :~~•mY(_~€(€€€C‡A2n(
lV!":R2:<=1283$3! 
:R2:0CI- ?8,(a ?h€•&@2!
:R2:,<=,0C6(
J ::!:R2:<=$C<Q 
D5=?ICx 2!22$2H?ICC@
3<( ;&  & !:R2: 2  r</  k2!:
J:!!! :l::N)<=$ &/&@28A
-<?ICC#/3C(
; @2&@2:R2:<=#"
_2=^^K
pW$U*&2*&,6!(6!,
@2!?@28i4+3#7#
#75D$ [(E 
V6!=3# &4/6 4! #
&4/6 $**^7(J&4!2
[V6!#(; .
&@2:R2:<=23=!CV&=(; 
&/&./3/2†/N./[
k2!:/Q !:R2&,6!T4(; ?IC
x 2 :R2:!: # :: <=&:R2: +
6!/H# Q(
p;28<=$C ?IC/
/D:x 2"4N :: O?^!,/#
^A[H,13C(k2!::::2:!!!:
 $* #&#!"D?(l ::^!2
2$2<• *!:?31,/3 #3X,/D:(

p;AH6!6(oR2:2# #
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
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Khóa luận tốt nghiệp Viện Đại Học Mở
<?@27+6!6(),
:R2: .23! &@2:R2: O $
?@2KC!2>C02C C N#?
>^,:R2:#$^>#^H!01?
IC&@2:R2:2"^2.#>0(
pJ?C!?@2K:R2:<=! /9
C6?1IC"^C?2D,@2(J&
@2:R2:X O 0#H2P?@2!: 
?ICx 2!: ?IC(
; @2 ::<Q!22$2H
/2†:R2<H#Hd,&!2 +<<
/X<=?ICx 2+,!22$2H(
oR2: @2,&<=?IC
./3(WV6.<=:R2: αpk2!:# βp
k2!:?IC27+/3?@28<:I! :#
2! :<Q$ /*‰ (y! :?IC
.αpk2!:#βpk2!:6! :<Q @2
23CU# * ?€(e!V.*^#5#7
<Q,& <(
A2Ydh€.?ICM :#I ! wx :
! .3CY(h]!V #".*^
 0$(
A2Ydde[!.6=6&"!Y€C5
6Lb6=_(\CM :m/ i2?I ! wx :n7!
e€€(€€€Co„!\CJ<]€€€€€CWŠ\€€€€€C
"!~€€€€€C(

)@2&@2:R2:<= 
<@2&&/&8A-(W>
#<=:R2:,##7 "? 
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
10
Khóa luận tốt nghiệp Viện Đại Học Mở
I!%2.68*<=(
; <#<=:R2:<=+
A2\€,&*‹\€(oR2:+"&L!#l::<=
Ti(A2YdhY<=:R2:k2!:,Z!!/!Ti1
 3  - P. (  oR2: y!:  l::  o:
l ::<= D5Cj2! /9V
*?J:!!! : (
; 3<D5<=:R2:!!
#!22$2<(.<=:R2: !!::2 
/ I:<:C.#,!6*&  !!:.
#7!*9<#162$20(
; /3>#@&@2:R2:
<=! ::!::!!! :2!: I< :<:A#74#
D?D5@(L34:R2:/[I# ! I
X#*:2<G<4A/ 2:! !!::!:N
:R2:<=@@#&/@/26<*4i
.!22H#A3i <(
; :R2:<Q?IC8A 3#7
j2AH8AA4<= 8A(J:R2:
<=#"2=^!:R2:,2!: ::>
!:!!! :#:2:!!! :(J-,C!"
<<'C=0.!2AD?<=8A(
W$%SD*@/H8A 3#7X
 #546,-^#"#A! 8

A(
),?!"2,$2A:R2:(
+&*‹Yd:R2:=<=/4! 6
( oR2: ; J2    <=  !2 6 #62  ! #&
0#&/9#62[#62*^D?(;/#62S
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
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Khóa luận tốt nghiệp Viện Đại Học Mở
2&*4#62=(J2 /! O<<(L3
4:R2:6<=/<<G1•:2
.#./l:m+<<!n(2344
[!V!&/jD5:R2:.
2,(F^<=#?ICl!! /[.
:R2::!!!:?IC<gIC/V}pklk#hw
kEJk+[!6*&uuuuuuFF(JQ#"
,*b7:#! &/ .?ICC
204U[#"!9:x: 
 2 :1.!#:N; A2V23
45&#/,:N)-^,
:R2:4#"X&#"
22{,* #,:R2:
6Q#"&/3#$*b750!:R2:{-
^$0 >/ !S#(
 #68:R2: !2# $H
.X68,:R2: 0+
 /  &!-(Z1#7:R2:X#X
D <$ 68D5&
,4X# $!S#*(
 :R2:326:R2:*?IC#"
D29#$:R2:><QP4: *b

7:68 :m:< !::n<=:R2„::
H!: "N
J &.#g jdd•*? A8<=,
:R2:#g1<$2A(;601680/?#$C-
#0&[:R2:21#,:R2:K
p;[:R2: KC ^0:R2:
[/j . &*&#[CPC
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Khóa luận tốt nghiệp Viện Đại Học Mở
0?I #&*&2I-C**A(W&23#
:R2: <=< !!2 ^
0::(
pZ&[C-:R2:K<=*b7<$!2/&[
H"C-:R2:/&[ ^CN.<=
 :?/&2 3,:R2:2=^!2
  ^ 0 >:R2: ^2"(
poR2:<= ^@ $H(
pZ? #2.K<=:R2:I!%C? #7!
<=#7!&(
I.3.2 Tình hình sản xuất và ứng dụng enzyme ở Việt Nam.
 F68#8<=:R2:1F2U2"7
-% #7*‹1!(J &F223
:R2: ?IC1D222"U<+!1D
292#X^2,.#6
8(oR2::+<</Xm;.WrZ* r3n
/ 2:!+<8mWrˆ4yr3np2!:! :2!:
 ::+##7m#@2n
&@2:R2:<=3C1"
!2!:,&<Q ?IC/.

! :.<NoR2:? :m ::n
A3/$,/C@"C2N
L34:R2:m,&7+" n<=3
 # C!?@2#<?@2(
J&@2 ::m<Q ?IC"C2&
/&Hn# R2/ mn!&@2i4+#7(
)?  !  &  @2  :R2:  6  =    A2  * ?  Y€€p]€€
C‡A2(;6#<=:R2: 1
"X1<$2A< #7 A2"*?A8
Khoa Công Nghệ Sinh Học Vũ Thị Hằng - 0604
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