Difference between revisions of "Part:BBa K176011"

Revision as of 06:34, 20 October 2009

pCon 1.00(J23100)->RBS+GFP+T

pCon 1.00(J23100)+I13504

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 705

MEASUREMENT

We measured five constitutive promoter family member ( J23100, J23101, J23103, J23107, J23109, J23115, )strength by ligated each with a 
GFP gene ( I13504) , and then moniter the OD600 and GFP expression.

data

J23100+GFP

COLONY NUMBER	T(min)	OD600	FLU(arbitrary)	ST.FLU(ustc_st.1)	FLU/OD600(arbitrary)	ST.FLU/OD600(ustc_st.1)
Ⅰ	0	0.071	132.07	527.9210137	1860.140845	7435.507235
Ⅰ	30	0.076	144.49	577.5672543	1901.184211	7599.569135
Ⅰ	60	0.101	157.6633333	630.2247805	1561.023102	6239.849312
Ⅰ	90	0.093	160.8866667	643.1093523	1729.964158	6915.154326
Ⅰ	120	0.116	176.33	704.8407083	1520.086207	6076.213003
Ⅰ	150	0.122666667	185.0633333	739.7503031	1508.668478	6030.573123
Ⅰ	180	0.153	197.8966667	791.0487535	1293.442266	5170.253291
Ⅱ	0	0.07	144.1	576.0083143	2058.571429	8228.690205
Ⅱ	30	0.075333333	136.1066667	544.0567081	1806.725664	7221.991701
Ⅱ	60	0.084333333	144.92	579.2860855	1718.418972	6869.004966
Ⅱ	90	0.092	157.4133333	629.22546	1711.014493	6839.407174
Ⅱ	120	0.098	164.73	658.4722389	1680.918367	6719.104478
Ⅱ	150	0.117	179.78	718.6313307	1536.581197	6142.148126
Ⅱ	180	0.135666667	194.6566667	778.0975603	1434.815725	5735.362853

For the details of the definition of ST.FLU and the unit ustc_st1, please click here.

plot

Protocol

1. Streak a plate of the strain which contain one of the parts listed in pSB1A3 .

2. Inoculate two 3ml cultures of supplemented M9 Medium and antibiotic( Ampicillin 0.1mg/ml) with single colony from the plate.

3. Cultures were grown in test tubes（BIO BASIC INC.12ml Polypropylene Round-bottom Culture Tubes With Graduations And Dual Cap Cat.No:TD444） for 16hrs at 37℃ with shaking at 200rpm.

4. Cultures were diluted 1:100 into 3ml fresh medium and grown for 3hrs.

5. Measure the fluorescence(SHIMDZU SPECTROFLUOROPHOTOMETER RF-5301PC, 250ul quartz cell path length 10mm,501 nm excitation,514 nm emission,1.5nm slit width) and absorbance (HITACHI UV-VIS spectrophotometer U-2810 ,200ul quartz cell,path length 10mm,600nm,1.5 nm slit width) every 30 minutes in the next 4hrs.

@@ Line 13: / Line 13: @@
 =='''MEASUREMENT'''==
-  We measured five '''constitutive promoter family member''' ( J23100, J23101, J23103, J23107, J23109, J23115, J23119 )strength by ligated each
+  We measured five '''constitutive promoter family member''' ( J23100, J23101, J23103, J23107, J23109, J23115, )strength by ligated each with a
- with a GFP gene ( I13504) , and then moniter the OD600 and GFP expression.
+ GFP gene ( I13504) , and then moniter the OD600 and GFP expression.
 ===data===
 J23100+GFP