scholarly journals Cloning and Expression Analysis of Mevalonate Kinase and Phosphomevalonate Kinase Genes Associated with MVA Pathway in Santalum Album

2020 ◽  
Author(s):  
Meiyun Niu ◽  
Yuping Xiong ◽  
Haifeng Yan ◽  
Xinhua Zhang ◽  
Yuan Li ◽  
...  
Keyword(s):  
Amino Acids ◽  
Sequence Comparison ◽  
Functional Complementation ◽  
Cloning And Expression ◽  
Santalum Album ◽  
Deficient Mutant ◽  
Cdna Synthesis ◽  
Bioinformatics Analyses ◽  
Mva Pathway ◽  
Young Leaves

Abstract Sandalwood is highly valued for its fragrant heartwood and its extracted oil. The major oil component santalols are terpenoids, which are biosynthesis through the MVA pathway. MK and PMK are the major enzymes on the MVA pathway. Little is known about the genes encoding MK and PMK in Santalum album on its expression regulation mechanism. The analysis of MK and PMK genes and their functions are important for the further study of the biosynthesis of santalol. These results will help to further study the role of MK and PMK genes in S. album santalol biosynthesis. The total RNA of sandalwood leaves was extracted, then the First-strand cDNA synthesis was obtained through the PrimeScript first-strand cDNA synthesis kit. Then sequence comparison and bioinformatics analyses of the genes homology of SaMK and SaPMK with MKs and PMKs, We also investigated subcellular localization of SaMK and SaPMK proteins. Its functional complementation of SaMK and SaPMK in yeast were also investigated. Atlast, MeJA was used to induce tissue-specific analysis and expression profiles of SaMK and SaPMK. The results showed that the full-length cDNA sequences of SaMK and SaPMK were 1409 bp and 1679 bp containing a 1381 bp open reading frame (ORF) encoding a polypeptide of 460 amino acids and a 1527 bp ORF encoding a polypeptide of 508 amino acids, respectively. Sequence comparison and bioinformatics analyses indicated that SaMK and SaPMK showed high homology with MKs and PMKs, respectively from other plant species. Further functional complementation of SaMK in an MK-deficient mutant yeast strain YMR208W and SaPMK in a PMK-deficient mutant yeast strainYMR220W confirmed that cloned SaMK and SaPMK cDNA encode a functional MK and PMK, respectively and mediated MVA biosynthesis in yeast. Tissue expression pattern analysis revealed that SaMK and SaPMK were constitutively expressed in all the tested tissues. SaMK was highly expressed in young leaves but least expressed in sapwood while SaPMK was highly expressed in roots and mature leaves, and least expressed in young leaves.

scholarly journals Cloning and expression analysis of mevalonate kinase and phosphomevalonate kinase genes associated with the MVA pathway in Santalum album

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Meiyun Niu ◽  
Yuping Xiong ◽  
Haifeng Yan ◽  
Xinhua Zhang ◽  
Yuan Li ◽  
...  
Keyword(s):  
Amino Acids ◽  
Yeast Strain ◽  
Functional Complementation ◽  
Cloning And Expression ◽  
Santalum Album ◽  
Mevalonate Kinase ◽  
Deficient Mutant ◽  
Mva Pathway ◽  
Young Leaves ◽  
Phosphomevalonate Kinase

AbstractSandalwood (Santalum album L.) is highly valued for its fragrant heartwood and extracted oil. Santalols, which are the main components of that oil, are terpenoids, and these are biosynthesized via the mevalonic acid (MVA) pathway. Mevalonate kinase (MK) and phosphomevalonate kinase (PMK) are key enzymes in the MVA pathway. Little is known about the genes that encode MK and PMK in S. album or the mechanism that regulates their expression. To isolate and identify the functional genes involved in santalol biosynthesis in S. album, an MK gene designated as SaMK, and a PMK gene designated as SaPMK, were cloned from S. album. The sequences of these genes were analyzed. A bioinformatics analysis was conducted to assess the homology of SaMK and SaPMK with MK and PMK genes from other plants. The subcellular localization of SaMK and SaPMK proteins was also investigated, as was the functional complementation of SaMK and SaPMK in yeast. Our results show that the full-length cDNA sequences of SaMK and SaPMK were 1409 bp and 1679 bp long, respectively. SaMK contained a 1381 bp open reading frame (ORF) encoding a polypeptide of 460 amino acids and SaPMK contained a 1527 bp ORF encoding a polypeptide of 508 amino acids. SaMK and SaPMK showed high homology with MK and PMK genes of other plant species. Functional complementation of SaMK in a MK-deficient mutant yeast strain YMR208W and SaPMK in a PMK-deficient mutant yeast strain YMR220W confirmed that cloned SaMK and SaPMK cDNA encode a functional MK and PMK, respectively, mediating MVA biosynthesis in yeast. An analysis of tissue expression patterns revealed that SaMK and SaPMK were constitutively expressed in all the tested tissues. SaMK was highly expressed in young leaves but weakly expressed in sapwood. SaPMK was highly expressed in roots and mature leaves, but weakly expressed in young leaves. Induction experiments with several elicitors showed that SaMK and SaPMK expression was upregulated by methyl jasmonate. These results will help to further study the role of MK and PMK genes during santalol biosynthesis in S. album.

scholarly journals Cloning and Expression Analysis of Two Kdm Lysine Demethylases in the Testes of Mature Yaks and Their Sterile Hybrids

Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 521
Author(s):  
Zhenhua Shen ◽  
Lin Huang ◽  
Suyu Jin ◽  
Yucai Zheng
Keyword(s):  
Amino Acids ◽  
Protein Expression ◽  
Male Sterility ◽  
Histone Methylation ◽  
Real Time Pcr ◽  
Cloning And Expression ◽  
Rt Pcr ◽  
Coding Sequences ◽  
Mrna And Protein Expression ◽  
Sterile Hybrids

The objective of this study was to explore the molecular mechanism for male sterility of yak hybrids based on two demethylases. Total RNA was extracted from the testes of adult yaks (n = 10) and yak hybrids (cattle–yaks, n = 10). The coding sequences (CDS) of two lysine demethylases (KDMs), KDM1A and KDM4B, were cloned by RT-PCR. The levels of KDM1A and KDM4B in yaks and cattle–yaks testes were detected using Real-time PCR and Western blotting for mRNA and protein, respectively. In addition, the histone methylation modifications of H3K36me3 and H3K27me3 were compared between testes of yaks and cattle–yaks using ELISA. The CDS of KDM1A and KDM4B were obtained from yak testes. The results showed that the CDS of KDM1A exhibited two variants: variant 1 has a CDS of 2622 bp, encoding 873 amino acids, while variant 2 has a CDS of 2562 bp, encoding 853 amino acids. The CDS of the KDM4B gene was 3351 bp in length, encoding 1116 amino acids. The mRNA and protein expression of KDM1A and KDM4B, as well as the level of H3K36me3, were dramatically decreased in the testes of cattle–yaks compared with yaks. The present results suggest that the male sterility of cattle–yaks might be associated with reduced histone methylation modifications.

Sequence and Expression of a Caffeic Acid O-Methyl Transferase Cdna Homologue in the Tropical Forage Legume Stylosanthes Humilis.

1995 ◽  
Vol 22 (3) ◽  
pp. 471 ◽  
Author(s):  
CL Mcintyre ◽  
AL Rae ◽  
MD Curtis ◽  
JM Manners
Keyword(s):  
Amino Acids ◽  
Caffeic Acid ◽  
Open Reading Frame ◽  
Stem Tissue ◽  
Forage Legume ◽  
Methyl Transferase ◽  
Reading Frame ◽  
Stylosanthes Humilis ◽  
Tropical Forage ◽  
Young Leaves

The isolation and characterisation of a cDNA encoding a caffeic acid 0-methyl transferase cDNA homologue (COMT) from Stylosanthes humilis are described. The clone is 1391 nucleotides in length, with an open reading frame encoding a predicted protein of 366 amino acids. Cluster analysis of the deduced amino acid sequence revealed extensive homology to other published O-methyl transferase sequences. Maximum levels of homology were seen with COMTs from alfalfa (87%) and aspen (84%). Southern analysis suggested that this enzyme is encoded by two genes in S. humilis. The mRNA is most strongly expressed in stem tissue, with intermediate levels of expression in young leaves and roots, and does not appear to be induced upon fungal infection or wounding.

Molecular cloning and expression of key enzymes for biosynthesis of cysteine and related secondary non-protein amino acids

1994 ◽  
Vol 38 (2-3) ◽  
pp. 153-158 ◽  
Author(s):  
Kazuki Saito ◽  
Naoko Miura ◽  
Mami Yamazaki ◽  
Kazuyo Tatsuguchi ◽  
Makoto Kurosawa ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Cloning ◽  
Cloning And Expression ◽  
Key Enzymes ◽  
Protein Amino Acids

Cloning and expression of a sesquiterpene synthase gene from Taiwania cryptomerioides

Holzforschung ◽  
2015 ◽  
Vol 69 (9) ◽  
pp. 1041-1048 ◽  
Author(s):  
Hui-Ling Hsieh ◽  
Li-Ting Ma ◽  
Sheng-Yang Wang ◽  
Fang-Hua Chu
Keyword(s):  
Major Product ◽  
Cloning And Expression ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Degenerate Primers ◽  
Sesquiterpene Synthase ◽  
Reading Frame ◽  
Spectrometry Analysis ◽  
Young Leaves ◽  
Taiwania Cryptomerioides

Abstract Taiwania (Taiwania cryptomerioides Hayata) is a conifer species native to Taiwan, which is known for several bioactive secondary metabolites extracted from it. In this study, a sesquiterpene synthase (TPS) gene isolated from Taiwania was in focus. First, a pair of degenerate primers was designed for reverse transcription-polymerase chain reaction based on the total RNA extracted from the leaves of a mature tree. A DNA fragment with the conserved region of TPS gene was obtained. After 5′- and 3′-end amplification, the full-length gene was obtained, which contains an open reading frame of 1791 bp and encodes a predicted molecular mass of 70.2-kDa protein. The gene was highly expressed in young leaves, female flowers, and cones. The expression in leaves was enhanced by salicylic acid. To identify the function of TPS, the recombinant protein from Escherichia coli (Migula) Castellani & Chalmers was incubated with farnesyl diphosphate. Gas chromatography/mass spectrometry analysis and retention time as well as mass spectrum matching with authentic standards revealed that the major product of TPS is sesquiterpene α-gurjunene. The gene was, therefore, designated as Tc-Gur.

scholarly journals Antidepressant Effect of Shaded White Leaf Tea Containing High Levels of Caffeine and Amino Acids

Molecules ◽  
2020 ◽  
Vol 25 (15) ◽  
pp. 3550
Author(s):  
Keiko Unno ◽  
Daisuke Furushima ◽  
Yuzuki Nomura ◽  
Hiroshi Yamada ◽  
Kazuaki Iguchi ◽  
...  
Keyword(s):  
Amino Acids ◽  
Green Tea ◽  
Pharmacy Practice ◽  
Antidepressant Effect ◽  
Stress Markers ◽  
White Leaf ◽  
Antistress Effect ◽  
Young Leaves ◽  
The Difference ◽  
Temperature Water

The young leaves of green tea become lighter in color than usual when protected from sunlight by a shading net for about two weeks while growing. These leaves are called “shaded white leaf tea” or SWLT. In the eluate of SWLT, the amount of amino acids (361 mg/L) was significantly higher than that in regular tea (53.5 mg/L). Since theanine and arginine, the first and second most abundant amino acids in SWLT, have significant antistress effects, we examined the antistress effect of SWLT on humans. SWLT or placebo green tea (3 g) was eluted with room-temperature water (500 mL). Participants consumed the tea for one week prior to pharmacy practice and continued for 10 days in the practice period. The state-trait anxiety inventory, an anxiety questionnaire, tended to be scored lower in the SWLT group than the placebo, but other stress markers showed no differences. The effect of the difference in SWLT components examined with mice showed that aspartic acid and asparagine, which are abundant in SWLT, counteracted the antistress effects of theanine and arginine. Large amounts of caffeine also interfered with SWLT’s antistress effect. Thus, SWLT, which is high in caffeine and amino acids, suppressed depressant behavior in mice.

Fluctuations of γ-aminobutyrate, γ-hydroxybutyrate, and related amino acids in Arabidopsis leaves as a function of the light–dark cycle, leaf age, and N stressEditorial decisions for this paper were made by Robert Ireland, Associate Editor, Canadian Journal of Botany.

2006 ◽  
Vol 84 (8) ◽  
pp. 1339-1346 ◽  
Author(s):  
Wendy L. Allan ◽  
Barry J. Shelp
Keyword(s):  
Amino Acids ◽  
Leaf Age ◽  
Associate Editor ◽  
Time Of Day ◽  
Multiple Sources ◽  
Dark Cycle ◽  
N Supply ◽  
Metabolic Role ◽  
Close Relationship ◽  
Young Leaves

To gain further insight into the metabolic role of γ-aminobutyrate (GABA), we determined the pool sizes of GABA and its catabolic products, alanine and γ-hydroxybutyrate (GHB), as well as key amino acids (Glu, Gln, Asp, Asn, Pro, Gly, Ser), in Arabidopsis leaves as a function of the light–dark cycle, leaf age (old versus young), and N stress (continuous versus interrupted N supply). Regardless of time of day and leaf age, there was a close relationship among Glu, GABA, and GHB when N was supplied continuously, indicating that GABA and GHB were probably derived exclusively from Glu and GABA, respectively. Ala was also closely linked to GABA in young leaves, but not in old leaves, a result consistent with the existence of multiple sources of Ala. The nature of the responses of GABA and GHB to an interrupted N supply depended on leaf age, and differed from responses exhibited by Glu, Gln, and Asn. Overall fluctuations in primary amino acids under both continuous and interrupted N supply, as well as those associated with photorespiration, aging, and stress, suggest that the old and young leaves chosen for study here function in Arabidopsis as source and sink leaves, respectively.

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