Question

You want to characterize the function of the Cellulose synthase-like C, CslC, gene that has been reported to be involved in the biosynthesis of xyloglucan in higher plants. Your lab specializes in working with the moss, Physcomitrella patens. The CslC gene has been identified in this moss through genome sequence analysis, but it has yet to be reported if the moss gene really makes xyloglucan. It has also been reported in higher plants, that the CslC protein complex has been localized to the golgi apparatus. You are tasked with finding out what this gene does in moss and also localizing it’s respective protein complex within the cell. Outline how you would go about this task and list each molecular method that would be used

Answer 1

According to the information, it is desired to know the function of a gene in the moss. Secondarily, the objective of this experiment is to understand the localization of the protein complex generated from this gene. These two tasks can be performed as below:

Experiment 1: To know the function of gene

In vitro synthesis of knock-out variety of the moss can be developed by using recombinant DNA technology. These recombinant mosses can be used to understand the failure in a specific biological function as compared to the normal one. The function lacking in the recombinant moss will be the one controlled by the gene of interest.

Experiment 2: To know the localization of the protein comlex

In vitro tagging of the DNA sequence for this gene can be performed. This DNA sequence will then be sequenced and amplified by PCR. The gene can be subjected to cell-free translation and then tagged with appropriate fluorescent tags. This tagged protein can be inserted back into the knock-out variety of moss and its individual cell can be monitored under real time fluorescent microscope. This will demonstrate the localization of the target protein in the cells.