The plasmid pGEX-3X-hHus1 containing GST-tagged hHus1 was obtained from Dr. A. E. Tomkinson at the University of New Mexico. GST fusions incorporating hHus1 deletion constructs were made by polymerase chain reaction (PCR) using primers listed in Table S1 in the supplementary material . The PCR products were digested with BamHI and SalI and ligated into the BamHI-XhoI-digested pGEX-4T-2 vector (GE Healthcare). The K136A and V137A mutants of the hHus1 gene were constructed by Quick Change site-directed mutagenesis (Stratagene) using pGEX-3X-hHus1 plasmid as a template and primers listed in Table S1 in the supplementary material .
Bamhi xhoi
Manufactured by GE Healthcare
Sourced in Japan
BamHI-XhoI is a restriction enzyme that recognizes and cleaves specific DNA sequences. It is commonly used in molecular biology and genetic engineering applications for DNA manipulation and analysis.
Automatically generated - may contain errors
3 protocols using bamhi xhoi
1
Generation of GST-tagged hHus1 Constructs
2
Cloning and Mutagenesis of Cyanobacterial PEPCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The region of the Synechocystis 6803 genome containing the ppc (sll0920, encoding SyPEPC) ORF was amplified by PCR using KOD plus neo polymerase and the primers 5′-GAAGGTCGTGGGATCATGAACTTGGCAGTTCCTG-3′ and 5′-GATGCGGCCGCTCGAGTCAACCAGTATTACGCATTC-3′. The amplified DNA fragments were cloned into the BamHI-XhoI site of pGEX5X-1 (GE Healthcare Japan, Tokyo, Japan) using an In-Fusion HD cloning kit (Takara Bio, Shiga, Japan). Site-directed mutagenesis was commercially performed by Takara Bio. For SyPEPC_E954K and SyPEPC_S967V, +2860–2862 and +2899–2901 from the start codon were changed from GAA to AAA and from TCT to GTG, respectively.
The region of the Anabaena 7120 genome containing the ppc (all4861, encoding AnPEPC) ORF was artificially synthesized and cloned into the BamHI-XhoI site of pGEX5X-1 by Takara Bio.
The region of the Anabaena 7120 genome containing the ppc (all4861, encoding AnPEPC) ORF was artificially synthesized and cloned into the BamHI-XhoI site of pGEX5X-1 by Takara Bio.
3
Expression Vectors for Ubiquitination Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The DNA fragments encoding human UBE2D1, UBE2N, and MMS2 were amplified by PCR from human HeLa Marathon-Ready cDNA (Clontech). The amplified DNA fragments encoding UBE2N and MMS2, or UBE2D1, or RFWD3 were ligated into the BamHI-SalI, or EcoRI-XhoI, or BamHI-XhoI sites of the pGEX-6P-1 vector (GE Healthcare), respectively. For the generation of the expression vector for the human RAD51-5KR mutant, the DNA fragment encoding the human RAD51 ORF carrying the 5KR mutation was ligated into the NdeI-BamHI sites of the pET-15b vector (Novagen). For the generation of the expression vector for the human RPA-5KR mutant, the DNA fragment encoding the human RPA2 ORF carrying the 5KR mutation was ligated into the p11d-tRPA vector (Henricksen et al., 1994) , using an In-Fusion HD Cloning Kit (Takara).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!