Difference between revisions of "Part:BBa I750016:Experience"

Latest revision as of 20:06, 21 October 2009

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_I750016

BBa_K190025 (Planning) constitutive promoter BBa_J23109 with GVP cluster

BBa_K190026 (Planning) constitutive promoter BBa_J23106 with GVP cluster

BBa_K190033 (Planning) metal (As) sensitive promoter BBa_K190015 with GVP cluster

BBa_K190034 (Planning) metal (Zn) sensitive promoter BBa_K190016 with GVP cluster

BBa_K190035 (Planning) metal (Cu) sensitive promoter BBa_K190017 with GVP cluster

BBa_K190036 (Planning) pLacI promoter BBa_R0010 with GVP cluster

User Reviews

UNIQd3b0b7c042ec9704-partinfo-00000006-QINU

•••••

Antiquity

This review comes from the old result system and indicates that this part worked in some test.

•••••

iGEM Groningen 2009

Part BBa_I750016 was one of our main focus points during our project. The part was originally submitted by Melbourne in 2007 with a very limited amount of information (short description and sequence mutations). On their site it was mentioned the cluster was difficult to use and ligation into a plasmid was hard, in contrast we found the cluster to be easily cut and ligated into different plasmids and behind several promotors. However we did not get results from PCR's that we did on vectors containg the GVP cluster. We used this part in combination with several biobricks for building our constructs e.g. BBa_K190033 and BBa_K190036. Pictures of our cells with induced gas vesicle formation confirmed the production of vesicles in the expected shape and size. Buoyancy tests showed indeed that the introduction of gas vesicle genes into E. coli led to increased buoyancy. Increased buoyancy means in our case that the cells sink slower (in saline). The cells don't float to the top layer of the tube. In addition, we characterized the part to improve the use in the future and added a lot of information on the registry.

UNIQd3b0b7c042ec9704-partinfo-0000000C-QINU Notes

21 May 2009

Phillip, Randy,

I'm writing with a question about BBa_I750016, Gas Vesicle polycistonic gene, as entered in the Registry of Standard Biological Parts:

https://parts.igem.org/Part:BBa_I750016

The long description states that the parts is ~5.7 kb long.

The sequence stored in the Registry is 6064 bp long

From examination, their appear to be 11 perfect direct repeats starting with "tctgcaaatta" in the DNA sequence stored in the Registry.

I couldn't find any annotated feature associated with such direct repeats, and thus wonder whether there is a mistake in the DNA sequence that the Registry is providing.

I have added this email to the parts experience page on the Registry.

Thank you, Drew

____

Drew Endy Stanford Bioengineering The BioBricks Foundation

2 September 2009

Drew,

we are currently working with BBa_I750016, Gas Vesicle polycistonic gene, as entered in the Registry of Standard Biological Parts:

https://parts.igem.org/Part:BBa_I750016

The plan is to remove the 10x 40bp (400bp total) repeat in gene gvpL, and thereby reduce the size of the biobrick to its original size of ~5.7 kb. In doing so, we also hope to remove the unwanted mutation next to the repeat section, and thus return the biobrick to its original form as in pNL29 without PstI and EcoRI sites.

As far as we could tell, there were no additional changes in the biobrick.

Besides the sequence, we are also trying to test the buoyancy of the brick, and characterize it in this way.

From iGEM Groningen,

Michael Verhoeven

UNIQd3b0b7c042ec9704-partinfo-0000000D-QINU UNIQd3b0b7c042ec9704-partinfo-0000000E-QINU

@@ Line 43: / Line 43: @@
 <I> iGEM Groningen 2009 </I>
 |width='60%' valign='top'|
-Part <partinfo>BBa_I750016</partinfo> was one of our main focus points during our project. The part was originally submitted by Melbourne in 2007 with a very limited amount of information (short description and sequence mutations). On their site it was mentioned the cluster was difficult to use and ligation into a plasmid was hard, in contrast we found the cluster to be easily cut and ligated into different plasmids and behind several promotors. However we did not get results from PCR's that we did on vectors containg the GVP cluster. We used this part in combination with several biobricks for building our constructs e.g. <partinfo>BBa_K190033</partinfo> and <partinfo>BBa_K190036</partinfo>. Pictures of our cells with induced gas vesicle formation confirmed the production of vesicles in the expected shape and size. Buoyancy tests showed indeed that the introduction of gas vesicle genes into ''E. coli'' led to increased buoyancy. Increased buoyancy means in our case that the cells sink slower (in saline). The cells don't float to the top layer of the tube. In addition, we characterized the part to improve the use in the future and added a lot of information on the registry.
+Part <partinfo>BBa_I750016</partinfo> was one of our main focus points during our project. The part was originally submitted by Melbourne in 2007 with a very limited amount of information (short description and sequence mutations). On their site it was mentioned the cluster was difficult to use and ligation into a plasmid was hard, in contrast we found the cluster to be easily cut and ligated into different plasmids and behind several promotors. However we did not get results from PCR's that we did on vectors containg the GVP cluster. We used this part in combination with several biobricks for building our constructs e.g. <partinfo>BBa_K190033</partinfo> and <partinfo>BBa_K190036</partinfo>. [[Team:Groningen/Project/Vesicle#Electron Microscopy|Pictures]] of our cells with induced gas vesicle formation confirmed the production of vesicles in the expected shape and size. [[Team:Groningen/Project/Vesicle#Buoyancy|Buoyancy tests]] showed indeed that the introduction of gas vesicle genes into ''E. coli'' led to increased buoyancy. Increased buoyancy means in our case that the cells sink slower (in saline). The cells don't float to the top layer of the tube. In addition, we characterized the part to improve the use in the future and added a lot of information on the registry.
 |}
 <!-- DON'T DELETE --><partinfo>BBa_B0034 EndReviews</partinfo>