Heterologous transcription of a polyketide synthase gene from the lichen forming fungi Usnea longissima (Article)

Research Journal of Biotechnology

Volume 11, Issue 10, 1 October 2016, Pages 16-21

Yi, W.^a, 

Neng, Z.^a, 

Xiaolong, Y.^a, 

Mei, H.^a, 

Jae-Seoun, H.^b, 

Yuming, Y.^a, 

Juan, W.^a 

 

^a  Laboratory of Forest Plant Cultivation and Utilization, Yunnan Academy of Forestry, Kunming Yunnan, China
^b  KORn Lichen Research Institute, Sunchon National University, Sunchon, South Korea 

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Abstract

Usnea longissima is traditional medicine in Asia and several polyketide synthase (PKS) genes responding bioactive compound were isolated from U. longissima. However, the intron splicing of PKS gene from U. longissima is not clear in Aspergilli. In this study, a PKS of U. longissima (UlPKS6) was ligated into vector pNQ-ArgB which contained the A. nidulans niiA promoter, the Neurospora crassaqa4 terminator, took the ampicillin resistance gene and arginine as a selectable marker, first and then UlPKS6 was transferred into Aspergillus nidulans by polyethylene glycol mediated protoplasts transformation method. The integrity and copy number of UlPKS6 in transformants were detected by long and accurate polymerase chain reaction (LA-PCR) and Southern blot respectively; Reverse transcription-polymerase chain reaction (RT-PCR) and sequence alignment were used to detect gene expression and intron splicing of UlPKS6 in transformants. Nine transformants were obtained with single full UlPKS6 gene DNA from 106 protoplast. Intron splicing of UlPKS6 in transformants is same with in lichen forming fungi. UlPKS6 was transferred into A. nidulans and expressed in A. nidluans successfully. The DNA from U. longissima or other lichen can be used in heterologous expression in A. nidulans directly.

Author keywords

Aspergillus nidulans; Heterologous expression; Polyketide synthase; Usnea longissima