Establishment of an Efficient Genetic Transformation System in

Overview

Journal J Fungi (Basel)

Date 2024 Feb 23

PMID 38392809

Authors

Xutong Wang

Mandi Wang

Jian Sun

Xiaolei Qu

Shixin Wang

Tingting Sun

Affiliations

Soon will be listed here.

Abstract

(1) Background: , a valuable medicinal fungus, has limited studies on its gene function due to the lack of a genetic transformation system. (2) Methods: This study aimed to establish an efficient -mediated transformation (ATMT) system for This study involved cloning the promoter (glyceraldehyde-3-phosphate dehydrogenase, ) of , reconstructing the transformation vector, optimizing the treatment of receptor tissues, and inventing a new method for screening positive transformants. (3) Results: The established ATMT system involved replacing the CaMV35S promoter of pCAMBIA-1301 with the promoter of to construct the pCAMBIA-SH- transformation vector. The vectors were then transferred to (EHA105) for infection. This study found that the transformation efficiency was higher in the infection using pCAMBIA-SH- vectors than using pCAMBIA-1301 vectors. The mycelia of were homogenized for 20 s and collected as the genetic transformation receptor. After 20 min of co-culture and 48 h of incubation in 15 mL PDL medium at 25 °C, new colonies grew. (4) Conclusions: These colonies were transferred to PDA medium (hygromycin 4 μg/mL, cefotaxime 300 μg/mL), and the transformation efficiency was determined to be 33.7% using PCR.

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