Sandeep K. Panda

8 Questions 9 Answers 0 Followers

Questions related from Sandeep K. Panda

How can I prevent recombination in plasmids without knocking out recA gene?

1. My strain doesn't have a recA knock out. 2. I use a 2 plasmid system for my work. One replicates through rolling circle and the other through theta replication. 2. I have toehold...

29 February 2024 8,009 2 View

How to predict the transcription start site in bacteria if the ORF is already known?

In some cases the -10 of the promoter is known, while in others it is not. In that case how do we predict the transcription start site (TSS)? I have tried using the popular tools available. They...

09 May 2023 3,404 6 View

Can putting repetitive sequences in separate plasmids prevent recombination events that would normally happen in a single plasmid system?

We know that putting repetitive sequences in a single plasmid may initiate recombination in those plasmids. If recombination happens, the sequences in between those repetitive sequences may get...

18 February 2023 1,053 10 View

What recombinase genes need to be knocked out in Lactococcus genome for preventing recombination in plasmids?

I am working with RNA switches for tuning few genes for production of metabolites in Lactococcus. I have observed recombination events in my plasmids when I culture the respective strains for...

09 February 2023 5,592 2 View

CRISPR sgRNA contains sequences for target (20nt), Cas9 handle (42nt) and S. pyogenes Terminator (41nt). Is there a need for host specific Terminator?

I am creating CRISPRi tool for Lactococcus lactis. Do I need to add an extra Lactococcus Terminator sequence at the end of the sgRNA or is the S. pyogenes Terminator enough?

28 April 2021 9,411 3 View

Is the gene expression level from single copy in the genome different from single copy of the gene in a single transformed plasmid (no genome copy)?

Question pertains to comparison of level of mRNA expression from single gene copy in the genome to single-copy-single-plasmid system. Kindly also explain the reasons for the same.

12 March 2021 3,822 6 View

How many different origin of replication (ori) sequences can I have in a plasmid?

Multiple ori sequences pertaining to different hosts are usually present in a plasmid. But how many different ones can I have, without causing problems? If problems do get created, what kind of...

26 November 2019 515 2 View

How can you predict the translation initiation rate from a sequence upsteam of start codon in a mRNA transcript?

What are the major factors that influence the above? Are there any good bioinformatic prediction tools for the same?

15 July 2019 3,463 3 View