Coding

Part:BBa_K3888008:Experience

Designed by: Xiaoxuan Cheng   Group: iGEM21_SJTang   (2021-10-01)
Revision as of 19:45, 20 October 2021 by RyanLei (Talk | contribs)


Applications of BBa_K3888008

Deletion Plasmid construction.png

Figure 1: Illustration of the general form of plasmid used to create marked knockout strains of R. palustris.

The origin of replication (ori), origin of transfer (oriT) and sacB (blue) are part of the delivery plasmid. The green sequence is inserted into the multiple cloning site of the delivery plasmid. The left and right fragments (green) are sequences amplified from the R. palustris genome and used in overlap extension PCR to generate a recombinant product containing (a deletion and a restriction site）. The restriction site is used to introduce the resistance cassette (purple). This interrupts the gene even if no deletion is present, and allows selection of successful transformants.
According to this model, we constructed UppE knockout plasmid below.
HupS Deletion Plasmid Map.png

Figure 2: Deletion plasmid of HupS constructed according to the method mentioned above.

User Reviews

UNIQea970afbdb4d0abf-partinfo-00000000-QINU
Reference: Rey, Federico E., Yasuhiro Oda, and Caroline S. Harwood. "Regulation of uptake hydrogenase and effects of hydrogen utilization on gene expression in Rhodopseudomonas palustris." Journal of bacteriology 188.17 (2006): 6143-6152. UNIQea970afbdb4d0abf-partinfo-00000001-QINU