Gene Cloning and Bioinformatics Analysis of Limonene-3-hydroxylase from Schizonepeta tenuifolia
10.13422/j.cnki.syfjx.20210112
- VernacularTitle:荆芥柠檬烯-3-羟化酶基因克隆及生物信息学分析
- Author:
Meng-jiao YIN
1
;
Shi-lin DAI
1
;
Pei-na ZHOU
1
;
Gu-yin LIN
1
;
Li-cheng LIU
1
;
Chan-chan LIU
1
;
Qi-nan WU
1
Author Information
1. College of Pharmacy，Collaborative Innovation Center of Chinese Medicinal Resources Industrialization，National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine，Nanjing University of Chinese Medicine，Nanjing 210023，China
- Publication Type:Research Article
- Keywords:
Schizonepeta tenuifolia;
limonene-3-hydroxylase;
monoterpene;
gene clone;
bioinformatics analysis
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(2):130-137
- CountryChina
- Language:Chinese
-
Abstract:
Objective:This paper aims to clone the cDNA sequence of limonene-3-hydroxylase（StL3OH） in Schizonepeta tenuifolia and analyze its sequence by bioinformatics. Method:Specific primers were designed based on sequences of StL3OH gene screened from transcriptome sequencing data of S. tenuifolia and the cDNA sequence of StL3OH gene was cloned by reverse transcription polymerase chain reaction （RT-PCR） and analyzed for its bioinformatics. Result:The StL3OH gene cDNA sequence length was 1 598 bp，containing a 1 497 bp long complete open reading frame which encoded 498 amino acids. StL3OH protein had a theoretical relative molecular mass of 56.40 kDa，with a hydrophilic and unstable nature. Bioinformatics analysis showed that StL3OH protein had no signal peptide but had a transmembrane domain which might be located in endoplasmic reticulum. Multiple sequence alignment and cluster analysis showed that the amino acid sequence of MsL3OH protein had a high similarity with StL3OH protein，both of which contained cytochrome P450 heme binding region，belonging to the D subfamily of cytochrome CYP71 family. Codon bias analysis showed that StL3OH gene preferred guanine/cytosine（G/C） ending codon，with 27 skewed codons， and Nicotiana benthamiana was proven to be the most suitable host for exogenous expression of StL3OH gene. Conclusion:The cDNA sequence of StL3OH gene was cloned from S. tenuifolia for the first time，which will provide a basis for further study on the structure and function of StL3OH protein and the regulation mechanism of StL3OH gene in the accumulation and biosynthesis of monoterpenes in S. tenuifolia.
