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暴马桑黄MVD基因cDNA全长克隆及表达特性分析(PDF/HTML)

《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

Issue:
2020年4期
Page:
79-85
Column:
研究论文
publishdate:
2020-09-01

Article Info:/Info

Title:
The cloning and expression analysis of mevalonate pyrophosphate decarboxylase gene cDNA sequence from Sanghuangporus baumii
Article ID:
1000-2006(2020)04-0079-07
Author(s):
LIU Zengcai1 SUN Tingting2 WANG Shixin1 MA Yisha1 WANG Xutong1 SUN Jian1 ZOU Li1
(1.College of Forestry, Northeast Forestry University, Harbin 150040, China; 2.Department of Food Engineering, Harbin University, Harbin 150086, China)
Keywords:
mevalonate pyrophosphate decarboxylase gene cloning bioinformatic analysis expression analysis
Classification number :
Q781
DOI:
10.3969/j.issn.1000-2006.201912007
Document Code:
A
Abstract:
Objective To clone and characterize a mevalonate pyrophosphate decarboxylase (MVD) gene involved in the triterpenoids biosynthesis pathway in Sanghuangporus baumii is to understand the regulation mechanism of the triterpenoids biosynthesis pathway. Method The characteristics of the MVD gene sequence were determined using a series of bioinformatic tools. The prokaryotic expression vector was constructed using homologous recombination. Moreover, qRT-PCR was performed to measure the MVD gene transcript level, and spectrophotometry was used to determine the content and variation of triterpenoids at different developmental stages of S. baumii. Result The sequence analysis showed that the MVD gene cDNA sequence length was 1 209 bp and encoded 402 amino acids. The molecular weight of the protein was predicted to be 43.43 ku, and it was named SbMVD (GenBank number MK977617). Amino acid sequence alignment revealed that the SbMVD protein shared the greatest homology with Ganoderma sinense and Grifola frondosa. SDS-PAGE electrophoresis showed that the target protein band was located at approximately 65 ku (including a 21 ku tag protein), which was consistent with the predicted protein. Furthermore, qRT-PCR and spectrophotometry results showed that the SbMVD gene transcript level and triterpenoids content increased before decreasing dynamically at different developmental stages. The variation trends were basically the same. Conclusion The cloning and analysis of the SbMVD gene indicated that it might play an important role in the triterpenoids biosynthesis pathway. Our results lay a foundation for further elucidating the triterpenoids biosynthesis function of the SbMVD gene in S. baumii.

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Last Update: 2020-08-13