Electroporation of Thalassiosira pseudonana v1

Abstract

An electroporation-mediated genetic transformation or the marine diatom Thalassiosira pseudonana was developed. Using a sorbitol-based buffer, T. pseudonana cells were successfully transformed with the Tpfcp/nat plasmid at an efficiency of 2820 per 10 8 cells. This represents a six-fold improvement compared with previously published methods.

Thalassiosira pseudonana (Cyclotella nana) (Hustedt) Hasle et Heimdal (Bacillariophyceae): A genetically tractable model organism for studying diatom biology, including biological silica formation

“…The episome is delivered with high efficiency (1500 transformants per 10 8 cells), does not become incorporated into the genome, and is maintained in the cell as long as the antibiotic is present (Karas et al, 2015). Recombinant DNA transfer by electroporation shows the highest transformation efficiency reported for T. pseudonana to date (3000 transformants per 10 8 cells; Bugge, 2015). For long‐term maintenance of transgenic T. pseudonana cell lines, a cryopreservation method was developed that shows post‐thaw viability levels of 70%–85% (Hipkin et al, 2014).…”

“…Techniques for incorporating recombinant DNA into a genome is a key feature for a model organism. For Thalassiosira pseudonana , this has so far been achieved by biolistic particle delivery (Poulsen et al, 2006), electroporation (Bugge, 2015), and bacterial conjugation (Karas et al, 2015). These methods rely on positive selection based on resistance to the antibiotics nourseothricin ( nat gene; Poulsen et al, 2006), zeocin ( ble gene; Scheffel et al, 2011), and blasticidin ( bsr gene; Heintze et al, 2022) with expression of the antibiotic resistance gene under control of endogenous promoters.…”

Natural Language Processing for Studying Consumer Journey: A Case Study of Sneaker Shoppers

“…Surveys can be time-consuming and expensive to conduct, and focus groups can be biased by the participants' self-reporting. 1 How to cite the artcle: Singh J.; Natural Language Processing for Studying Consumer Journey: A Case Study of Sneaker Shoppers, IJRST, Apr-Jun 2020, Vol 10, Issue 2, 98-101, DOI: http://doi.org/10.37648/ijrst.v10i02.009 NLP and data mining offer a new way to study consumer behavior by analyzing large amounts of unstructured data, such as social media posts. NLP techniques can be used to extract meaning from text data, and data mining techniques can be used to identify patterns and trends in the data.…”