Cloning, expression and bioinformatics analysis of BrrLOX7 gene in turnip

doi:10.3969/j.issn.1004-1524.2023.04.10

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (4): 831-840.DOI: 10.3969/j.issn.1004-1524.2023.04.10

• Horticultural Science • Previous Articles Next Articles

Cloning, expression and bioinformatics analysis of BrrLOX7 gene in turnip

YAN Cunyao(), JIA Kai, YAN Huizhuan(), GAO Jie

College of Horticulture, Xinjiang Agricultural University, Urumqi 830052, China

Received:2021-12-13 Online:2023-04-25 Published:2023-05-05

Abstract

Abstract:

Lipoxygenase (EC 1.13.11.12; LOX) is a class of fatty acid dioxygenases composed of non-heme iron, which plays an important role in plant growth and development and environmental stress response. In this study, the BrrLOX7 gene was cloned from turnip using homologous cloning techniques. The sequence of BrrLOX7 gene was analyzed by bioinformatics. The expression pattern of BrrLOX7 gene under abiotic stress was analyzed using fluorescence quantitative PCR. The results showed that the CDS length of cloned BrrLOX7 gene was 2 715 bp, it encoded 904 amino acids, protein encoded by BrrLOX7 gene had a relative molecular weight of 103.10 ku and had no transmembrane structure or signal peptide. Subcellular localization prediction showed that the protein encoded by this gene was located in chloroplasts, and the protein had the closest sequence homology and genetic distance to Chinese cabbage and radish. Expression patterns under different stresses showed that the BrrLOX7 gene was involved in response to four stresses (salt stress, cold stress, heat stress and drought stress). The results of this study will provide a basis for functional studies of the BrrLOX7 gene.

Key words: turnip, lipoxygenase, gene cloning, bioinformatics analysis, expression analysis

CLC Number:

S631.3

YAN Cunyao, JIA Kai, YAN Huizhuan, GAO Jie. Cloning, expression and bioinformatics analysis of BrrLOX7 gene in turnip[J]. Acta Agriculturae Zhejiangensis, 2023, 35(4): 831-840.

Figures/Tables 11

Fig.1 RT-PCR of BrrLOX7 M, DL5000 marker; 1 and 2 indicate PCR amplification products of the BrrLOX7 gene.

Table 1 Amino acid composition of the protein encoded by BrrLOX7 gene

氨基酸的种类 Types of amino acid	氨基酸的数目 Number of amino acids	占氨基酸总量的百分比 Percent of total amino acids/%	氨基酸的种类 Types of amino acid	氨基酸的数目 Number of amino acids	占氨基酸总量的百分比 Percent of total amino acids/%
苯丙氨酸Phe	34	3.76	脯氨酸Pro	62	6.86
亮氨酸Leu	89	9.85	苏氨酸Thr	45	4.98
异亮氨酸Ile	47	5.20	丙氨酸Ala	51	5.64
甲硫氨酸Met	16	1.77	酪氨酸Tyr	34	3.76
缬氨酸Val	53	5.86	组氨酸His	22	2.43
半胱氨酸Cys	12	1.33	谷氨酰胺Gln	25	2.77
色氨酸Trp	21	2.32	天冬氨酸Asn	30	3.32
精氨酸Arg	49	5.42	赖氨酸Lys	63	6.97
丝氨酸Ser	60	6.64	天冬氨酸Asp	61	6.75
甘氨酸Gly	55	6.08	谷氨酸Glu	75	8.30

Fig.2 Phosphorylation site prediction of BrrLOX7 protein

Fig.3 BrrLOX7 gene encodes a protein with hydrophilicity

Fig.4 Secondary structure of BrrLOX7 protein Blue for α-helix; red for extended chain; green for β-turn; purple for irregularly curled.

Fig.5 Tertiary structure of BrrLOX7 protein

Fig.6 Structural domain prediction of the BrrLOX7 protein sequence

Fig.7 Comparison of amino acid sequence of BrrLOX7 protein in turnip and other plants

Fig.8 Phylogenetic tree of turnip BrrLOX7 protein and other plant-related protein sequences

Fig.9 Prediction of cis-acting elements of the BrrLOX7 gene promoter in turnip

Fig.10 Expression of the BrrLOX7 gene in turnip under abiotic stress

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Cloning, expression and bioinformatics analysis of BrrLOX7 gene in turnip

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