Team:KIT-Kyoto/Parts

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Latest revision as of 03:16, 27 October 2010

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We used these parts for making our original biobrick parts

1.We obtained these parts from 2010 iGEM kit

Name	Part type	Resistance	Insert	Vector	Contents
<partinfo>pSB3k3</partinfo>(<partinfo>BBa_J04450</partinfo>)	Plasmid backbone	Kanamycin	738bp	2750bp	promoter(LacI) +RBS+mRFP+T+T
<partinfo>pSB6A1</partinfo>(<partinfo>BBa_J04450</partinfo>)	Plasmid backbone	Ampicillin	738bp	4032bp	promoter(LacI) +RBS+mRFP+T+T
<partinfo>pSB1C3</partinfo>(<partinfo>BBa_J04450</partinfo>)	Plasmid backbone	Chloramphenicol	1069bp	2072bp	promoter(LacI) +RBS+mRFP+T+T
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_I3522</partinfo>)	Composite	Ampicillin	937bp	2079bp	promoter(TetR)+RBS+GFP+T+T
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_R0040</partinfo>)	Regulatory	Ampicillin	54bp	2079bp	promoter(TetR)
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_I732019</partinfo>)	Regulatory	Ampicillin	3230bp	2079bp	RBS+LacZ+T+T
<partinfo>pSB1AK3</partinfo>(<partinfo>BBa_I732950</partinfo>)	Protein domain	Kanamycin	3230bp	2079bp	RBS+LacZ+T+T
<partinfo>pSB4A5</partinfo>(<partinfo>BBa_K193602</partinfo>)	Composite	Ampicillin	1896bp	3395bp	promoter(LacIQ)+RBS+melA
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_E0240</partinfo>)	Protein domain	Ampicillin	883bp	2079bp	RBS+GFP+T+T
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_I13507</partinfo>)	Protein domain	Ampicillin	937bp	2079bp	RBS+RFP+T+T
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_E0420</partinfo>)	Protein domain	Ampicillin	878bp	2079bp	RBS+CFP+T+T
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_E0430</partinfo>)	Protein domain	Ampicillin	878bp	2079bp	RBS+YFP+T+T
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_I13600</partinfo>)	Composite	Ampicillin	940bp	2079bp	promoter(TetR)+RBS+CFP+T+T
<partinfo>pSB1A2</partinfo>(<partinfo>BBa_J22005</partinfo>)	Composite	Ampicillin	2079bp	2623bp	promoter(TetR)+RBS+YFP+T+T

2.We obtained this part directly from iGEM HQ

We added new information to the BioBrick Part (pSB6A1) shown below that was originally developed by the 09’Tokyo-Tech group. This BioBrick Part having the backbone of pSB6, a low copy plasmid was designed to express GFP in E. coli. Generally low copy plasmid is more efficient in protein expression in compared with high copy plasmid such as pSB1. However no quantitative data on the efficiency of GFP expression with this plasmid was described in any site of iGEM. We therefore measured intensities of fluorescence of GFP protein produced in E. coli carrying the pSB6A1 and compared with that carrying the pSB1A2. The data clearly showed that expression level of GFP is much higher in E. coli carrying pSB6A1 than that carrying pSB1A2. Thus we decided to use pSB6 vector to measure activities of various promoters responsible to H₂O₂. We thank the 09’Tokyo-Tech group who has originally developed this part.This new information would be useful for the other iGEM teams in future.

>>About pSB1 vs pSB6

Name	Part Type	Resistance	Insert	Vector
<partinfo>pSB6A1</partinfo>(<partinfo>BBa_K121013</partinfo>)	Protein domain	Ampicillin	883bp	4022bp	RBS+GFP+T+T

We designed and constructed these parts originally.

We cloned the following inserts into the vector, pSB6A1. Out of them we cloned the two inserts(BBa_K362005, BBa_K362007) into the pSB1C3 to send to the iGEM headquarter.

<groupparts>iGEM010 KIT-Kyoto</groupparts>

@@ Line 37: / Line 37: @@
 |<partinfo>pSB1AK3</partinfo>(<partinfo>BBa_I732950</partinfo>)||Protein domain||Kanamycin||3230bp||2079bp||RBS+LacZ+T+T
 |- style="background-color:#eaf4fc;"
-|<partinfo>pSB4A5</partinfo>(<partinfo>BBa_K193602</partinfo>)||Composite||Ampicillin||1896bp||3395bp||promoter(Lac)+RBS+melA+T+T|
+|<partinfo>pSB4A5</partinfo>(<partinfo>BBa_K193602</partinfo>)||Composite||Ampicillin||1896bp||3395bp||promoter(LacIQ)+RBS+melA
 |- style="background-color:#eaf4fc;"
 |<partinfo>pSB1A2</partinfo>(<partinfo>BBa_E0240</partinfo>)||Protein domain||Ampicillin||883bp||2079bp||RBS+GFP+T+T
@@ Line 54: / Line 54: @@
 === 2.We obtained this part directly from iGEM HQ ===
-:We added new information to the BioBrick Part (pSB6A1) shown below that was originally developed by the 09’Tokyo-Tech group. This BioBrick Part having the backbone of pSB6, a low copy plasmid was designed to express GFP in <I>E. coli</I>. Generally low copy plasmid is more efficient in protein expression in compared with high copy plasmid such as pSB1. However no quantitative data on the efficiency of GFP expression with this plasmid was described in any site of iGEM. We therefore measured intensities of fluorescence of GFP protein produced in <I>E. coli</I> carrying the pSB6A1 and compared with that carrying the pSB1A2. The data clearly showed that expression level of GFP is much higher in E. coli carrying pSB6A1 than that carrying pSB1A2. Thus we decided to use pSB6 vector to measure activities of various promoters responsible to H<sub>2</sub>O<sub>2</sub>. We thank the 09’Tokyo-Tech group who has originally developed this part.<BR>
+:We added new information to the BioBrick Part (pSB6A1) shown below that was originally developed by the 09’Tokyo-Tech group. This BioBrick Part having the backbone of pSB6, a low copy plasmid was designed to express GFP in <I>E. coli</I>. Generally low copy plasmid is more efficient in protein expression in compared with high copy plasmid such as pSB1. However no quantitative data on the efficiency of GFP expression with this plasmid was described in any site of iGEM. We therefore measured intensities of fluorescence of GFP protein produced in <I>E. coli</I> carrying the pSB6A1 and compared with that carrying the pSB1A2. The data clearly showed that expression level of GFP is much higher in E. coli carrying pSB6A1 than that carrying pSB1A2. Thus we decided to use pSB6 vector to measure activities of various promoters responsible to H<sub>2</sub>O<sub>2</sub>. We thank the 09’Tokyo-Tech group who has originally developed this part.This new information would be useful for the other iGEM teams in future.
+<BR>
 :[https://2010.igem.org/Team:KIT-Kyoto/Project/Abstract#2._Compare_the_performances_of_the_high_copy_vector_and_low_copy_vector >>About pSB1 vs pSB6]