ja, gene cloning and DNA analysis from T.A.Brown is my basic book for my life science cource. Let me tell u all in more details about what i'm doing. We' re now doing a project about L.lactis fermentation. All we have to do is to look for a gene from different organisms and transfer it to L.Lactis to control its' metabolic flux since L.lactis has the potential to produce a variety of end metabolic during sugar fermentation.
We hope to modify the metabolic flux in L. lactis by using metabolic engineering on the level of:
- NADH oxidase (key cofactor)
- Enhancement of ALS gene
- Inactivation of LDH gene
So now we have to look for some als or NADH oxidase (Nox) gene from the orther organisms and insert to vector pNZ8048 in L.lactis and then transform it to L.lactis bacteria to do fermentation. That's why we need to do so many steps such as cultivation of the cell culture, isolation, purification, PCR, topoTA, digestion, ligation, transformation, fermentation, etc.
So i need to have some book to give us more information about the protocols, the troubles we're gonna face with during the process, why we have to do this step, not the other steps, and so on. And i want to have some books with questions and answers around this subject for instance the book will give us the questions and answers for: how can u design the primers for the sequences, why you need to do topoTA, or what are the functions of the substrates u use, etc.
That's what i wanna know, plz give me some advice because my teachers really like asking those questions during exams, presentation, etc.
