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1.
Cell Death Differ ; 2021 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-34497381

RESUMO

Aged bone marrow mesenchymal stem cells (BMSCs) exhibit aberrant self-renewal and lineage specification, which contribute to imbalanced bone-fat and progressive bone loss. In addition to known master regulators of lineage commitment, it is crucial to identify pivotal switches governing the specific differentiation fate of aged BMSCs. Here, we profiled differences in epigenetic regulation between adipogenesis and osteogenesis and identified super-enhancer associated lncRNA nuclear-enriched abundant transcript 1 (NEAT1) as a key bone-fat switch in aged BMSCs. We validated that NEAT1 with high enhancer activity was transcriptionally activated by ATF2 and directed aged BMSCs to a greater propensity to differentiate toward adipocytes than osteoblasts by mediating mitochondrial function. Furthermore, we confirmed NEAT1 as a protein-binding scaffold in which phosphorylation modification of SOX2 Ser249/250 by CDK2 impaired SOX2/OCT4 complex stability and dysregulated downstream transcription networks of pluripotency maintenance. In addition, by sponging miR-27b-3p, NEAT1 upregulated BNIP3L, BMP2K, and PPARG expression to shape mitochondrial function and osteogenic/adipogenic differentiation commitment, respectively. In extracellular communication, NEAT1 promoted CSF1 secretion from aged BMSCs and then strengthened osteoclastic differentiation by extracellular vesicle delivery. Notably, Neat1 small interfering RNA delivery induced increased bone mass in aged mice and decreased fat accumulation in the bone marrow. These findings suggest that NEAT1 regulates the lineage fates of BMSCs by orchestrating mitochondrial function and pluripotency maintenance, and might be a potential therapeutic target for skeletal aging.

2.
Nat Commun ; 12(1): 4452, 2021 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-34294713

RESUMO

Atherosclerosis-associated cardiovascular disease is one of the main causes of death and disability among patients with diabetes mellitus. However, little is known about the impact of S-nitrosylation in diabetes-accelerated atherosclerosis. Here, we show increased levels of S-nitrosylation of guanine nucleotide-binding protein G(i) subunit alpha-2 (SNO-GNAI2) at Cysteine 66 in coronary artery samples from diabetic patients with atherosclerosis, consistently with results from mice. Mechanistically, SNO-GNAI2 acted by coupling with CXCR5 to dephosphorylate the Hippo pathway kinase LATS1, thereby leading to nuclear translocation of YAP and promoting an inflammatory response in endothelial cells. Furthermore, Cys-mutant GNAI2 refractory to S-nitrosylation abrogated GNAI2-CXCR5 coupling, alleviated atherosclerosis in diabetic mice, restored Hippo activity, and reduced endothelial inflammation. In addition, we showed that melatonin treatment restored endothelial function and protected against diabetes-accelerated atherosclerosis by preventing GNAI2 S-nitrosylation. In conclusion, SNO-GNAI2 drives diabetes-accelerated atherosclerosis by coupling with CXCR5 and activating YAP-dependent endothelial inflammation, and reducing SNO-GNAI2 is an efficient strategy for alleviating diabetes-accelerated atherosclerosis.


Assuntos
Aterosclerose/etiologia , Aterosclerose/metabolismo , Angiopatias Diabéticas/etiologia , Angiopatias Diabéticas/metabolismo , Subunidade alfa Gi2 de Proteína de Ligação ao GTP/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Células Cultivadas , Cisteína/química , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/metabolismo , Subunidade alfa Gi2 de Proteína de Ligação ao GTP/química , Subunidade alfa Gi2 de Proteína de Ligação ao GTP/genética , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Melatonina/farmacologia , Camundongos , Camundongos Knockout , Mutagênese Sítio-Dirigida , Óxido Nítrico Sintase Tipo II/metabolismo , Compostos Nitrosos/química , Compostos Nitrosos/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Receptores CXCR5/deficiência , Receptores CXCR5/genética , Receptores CXCR5/metabolismo , Fatores de Transcrição/metabolismo
3.
Front Pharmacol ; 12: 688073, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34262457

RESUMO

Insufficient pancreatic ß-cell or insulin-producing ß-cell are implicated in all types of diabetes mellitus. Our previous studies showed bee pollen polysaccharide RBPP-P improves insulin resistance in type 2 diabetic mice by inhibiting liver fat deposition. However, its potential of regulating ß-cell function and integrity is not fully known. Herein, we observed that ß-cell proliferation (n = 10), insulin synthesis (n = 5, p = 0.01684) and insulin incretion (n = 5, p = 0.02115) were intensely activated in MIN6 cells when treatment with RBPP-P. In alloxan-induced diabetic mice, oral administration of RBPP-P (n = 10) effectively decreased the blood glucose (p = 0.0326), drink intake (p < 0.001) and urine (p < 0.001). It directly stimulated phosphorylation of p38 (p = 0.00439), ERK (p = 0.02951) and AKT (p = 0.0072) to maintain the islet function and mass. Thus, our data suggest that RBPP-P is a natural compound to regulate ß-cell proliferation and function, indicating it might have therapeutic potential against type 1 diabetes.

4.
Front Cell Dev Biol ; 9: 675411, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34124061

RESUMO

It has been well established that leukemia inhibitory factor (LIF) is essential for maintaining naïve pluripotency of embryonic stem cells (ESCs). Oncostatin M (OSM) is a member of the IL-6 family of cytokines which share gp130 as a receptor subunit, and the OSM-gp130 complex can recruit either LIF receptor ß or OSM receptor ß. Here we show that OSM can completely replace LIF to maintain naïve pluripotency of ESCs. Mouse ESCs (mESCs) cultured in the presence of LIF or OSM not only express pluripotency genes at similar levels but also exhibit the same developmental pluripotency as evidenced by the generation of germline competent chimeras, supporting previous findings. Moreover, we demonstrate by tetraploid embryo complementation assay, the most stringent functional test of authentic pluripotency that mESCs cultured in OSM produce viable all-ESC pups. Furthermore, telomere length and telomerase activity, which are also crucial for unlimited self-renewal and genomic stability of mESCs, do not differ in mESCs cultured under OSM or LIF. The transcriptome of mESCs cultured in OSM overall is very similar to that of LIF, and OSM activates Stat3 signaling pathway, like LIF. Additionally, OSM upregulates pentose and glucuronate interconversion, ascorbate and aldarate metabolism, and steroid and retinol metabolic pathways. Although the significance of these pathways remains to be determined, our data shows that OSM can maintain naïve pluripotent stem cells in the absence of LIF.

5.
Am J Transl Res ; 13(4): 2555-2562, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34017414

RESUMO

OBJECTIVE: This study aimed to focus on the atlantoaxial pedicle screw placement and evaluate the effects of anatomical data and degree of surgical exposure of the atlantoaxial pedicle screw trajectory determined using spiral CT on the reference sites for pedicle screw insertion and various parameters in clinical application. METHOD: The data of CT scan of cervical spine from individuals treated in our hospital were selected. Various anatomical parameters of the atlantoaxial pedicle screw trajectory were measured through multiplanar reconstruction (MPR) technology. Anatomical data and degree of surgical exposure of the atlantoaxial pedicle screw trajectory were obtained through spiral CT. Vernier calipers with least count of 0.01 mm was selected and the least count of the protractor was 0.2°. Prism 8.0 was adopted for graphical data analysis. The anatomical data of atlas of the local population were established. The measurement technique for pedicle screw trajectory and the method for pedicle screw insertion were mastered. RESULTS: The results indicated that the intraoperative blood loss was between 30-280 ml with no case of excessive blood loss. Follow-up studies 10-18 months after the operations indicated stability of upper cervical spine without adverse conditions. The width of the pedicle screw was 13-24 mm, and the maximal inclination angle of the horizontal for the insertion was 17-21°. CONCLUSION: The atlantoaxial pedicle screw placement was an effective surgical treatment for stabilizing the upper cervical spine. The measurement data of the atlantoaxial pedicle screw path was obtained through spiral CT and the surgical placement of pedicle screw was guided through individual data. Postoperative CT scan was adopted to evaluate the accuracy of pedicle screw placement, record the occurrence of secondary injuries, improve the stability of clinical applications, and reduce the risks for patients.

6.
Acta Pharmacol Sin ; 2021 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-34011968

RESUMO

Cardiac fibrosis (CF) is an irreversible pathological process that occurs in almost all kinds of cardiovascular diseases. Phosphorylation-dependent activation of c-Jun N-terminal kinase (JNK) induces cardiac fibrosis. However, whether S-nitrosylation of JNK mediates cardiac fibrosis remains an open question. A biotin-switch assay confirmed that S-nitrosylation of JNK (SNO-JNK) increased significantly in the heart tissues of hypertrophic patients, transverse aortic constriction (TAC) mice, spontaneously hypertensive rats (SHRs), and neonatal rat cardiac fibroblasts (NRCFs) stimulated with angiotensin II (Ang II). Site to site substitution of alanine for cysteine in JNK was applied to determine the S-nitrosylated site. S-Nitrosylation occurred at both Cys116 and Cys163 and substitution of alanine for cysteine 116 and cysteine 163 (C116/163A) inhibited Ang II-induced myofibroblast transformation. We further confirmed that the source of S-nitrosylation was inducible nitric oxide synthase (iNOS). 1400 W, an inhibitor of iNOS, abrogated the profibrotic effects of Ang II in NRCFs. Mechanistically, SNO-JNK facilitated the nuclear translocation of JNK, increased the phosphorylation of c-Jun, and induced the transcriptional activity of AP-1 as determined by chromatin immunoprecipitation and EMSA. Finally, WT and iNOS-/- mice were subjected to TAC and iNOS knockout reduced SNO-JNK and alleviated cardiac fibrosis. Our findings demonstrate an alternative mechanism by which iNOS-induced SNO-JNK increases JNK pathway activity and accelerates cardiac fibrosis. Targeting SNO-JNK might be a novel therapeutic strategy against cardiac fibrosis.

7.
Medicine (Baltimore) ; 100(6): e24627, 2021 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-33578576

RESUMO

RATIONALE: Steroid-resistant nephrotic syndrome (SRNS) is a special kidney disease. SRNS is characterized by steroid-resistant, clinical variability, and genetic heterogeneity. Patients with SRNS often may eventually need renal transplantation. PATIENT CONCERNS: A 10-month-old Chinese male infant presented with oliguria, renal dysfunction, hypertension, and anemia. DIAGNOSES: Combined with clinical manifestations, laboratory testing and sequencing results, the patient was diagnosed as SRNS. INTERVENTIONS: Combined intravenous methylprednisolone and cefoperazone sulbactam did not improve the patient's condition. Thus, SRNS associated with hereditary nephrotic syndrome was strongly suspected. Genetic testing for hereditary renal disease of the patient revealed 2 novel heterozygous mutations in the Nucleoporin 93 (NUP93) gene, which were predicted pathogenic and harmful by bioinformatic softwares of SIFT, PolyPhen_2 and REVEL. OUTCOMES: As general physical health deterioration and renal dysfunction, the patient died of a severe infection. LESSONS: The novel NUP93 heterozygous mutations identified in the current study broadened the genetic spectrum of SRNS and further deepened our insight into pathogenic mutations of NUP93 to improve disease diagnosis.


Assuntos
Síndrome Nefrótica/diagnóstico , Antibacterianos/administração & dosagem , Antibacterianos/uso terapêutico , Cefoperazona/administração & dosagem , Cefoperazona/uso terapêutico , Evolução Fatal , Aconselhamento Genético , Glucocorticoides/administração & dosagem , Glucocorticoides/uso terapêutico , Humanos , Lactente , Masculino , Metilprednisolona/administração & dosagem , Metilprednisolona/uso terapêutico , Síndrome Nefrótica/tratamento farmacológico , Síndrome Nefrótica/genética
8.
J Cell Mol Med ; 25(1): 345-357, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33320446

RESUMO

In this study, we investigated the protective effects of gastrodin (Gas) against homocysteine-induced human umbilical vein endothelial cell (HUVEC) injury and the role of the phosphoinositide 3-kinase (PI3K)/threonine kinase 1 (Akt)/endothelial nitric oxide synthase (eNOS) and NF-E2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathways. We stimulated cells with homocysteine (1 mmol/L, 24 hours) and tested the effects of gastrodin (200-800 µg/mL) on cell viability and the production of malondialdehyde (MDA), lactate dehydrogenase (LDH) and reactive oxygen species (ROS). Then, Nrf2 distribution in the cytoplasm and nucleus as well as the expression of enzymes downstream of Nrf2 was determined. Furthermore, we analysed the expression of bax, bcl-2 and cleaved caspase3, and assessed the involvement of the PI3K/Akt/eNOS pathway by Western blots. Finally, we tested the vasoactive effect of gastrodin in thoracic aortic rings. The results showed that gastrodin decreased MDA, LDH and ROS production and increased cell viability, NO production and relaxation of thoracic aortic rings. Moreover, the protective effects of Gas on NO production and relaxation of thoracic aortic rings were blocked by L-NAME but enhanced by Cav-1 knockdown, and MK-2206 treatment abolished the effect of Gas on the ROS. In addition, treatment with gastrodin increased Nrf2 nuclear translocation, thus enhancing the expression of downstream enzymes. Finally, gastrodin increased the expression of PI3K, p-Akt, and eNOS and decreased Cav-1 protein expression. In conclusion, our study suggested that gastrodin may protect HUVECs from homocysteine-induced injury, and the PI3K/Akt/eNOS and Nrf2/ARE pathways may be responsible for the efficacy of gastrodin.


Assuntos
Álcoois Benzílicos/uso terapêutico , Glucosídeos/uso terapêutico , Homocisteína/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Estresse Oxidativo/efeitos dos fármacos
9.
Front Surg ; 7: 591522, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33330608

RESUMO

Purpose: Transoral endoscopic thyroidectomy via vestibular approach (TOETVA), with its excellent cosmetic effect, has become increasingly popular worldwide. Nonetheless, anatomic obstacles have limited its development to a certain extent. Here, we present our preliminary outcomes of transoral endoscopic thyroidectomy via submental and vestibular approach (TOETSMVA), which can overcome those limitations. Methods: From November 2019 to March 2020, we performed TOETSMVA in 21 consecutive patients with thyroid carcinoma at Zhongshan Hospital, Xiamen University. A 1.5-cm lateral incision was made at two fingers below the mandible; two 5-mm incisions were made in the vestibule near the first molars; TOETSMVA was completed through these incisions. The demographic data and surgical outcomes of the patients were retrospectively reviewed. Results: Twenty-one patients with a mean age of 37.5 ± 10.4 years were incorporated into this study. Fourteen patients had papillary thyroid micro-carcinomas, two had papillary thyroid carcinomas, and five had benign nodules. Eight patients had lymph node metastases. All surgeries were performed successfully without conversion to open thyroidectomy. The mean operation time was 138.8 ± 33.2 min; the average hospital stay was 3.3 ± 0.8 days. No patients developed cutaneous paralysis in the midline chin region. Transient recurrent laryngeal nerve paralysis was observed in one patient. There was no evidence of postoperative bleeding, infection, tetany, or other complications. Conclusion: TOETSMVA was shown to be a safe and advisable alternative for selected patients. This approach can overcome the limitations of TOETVA without sacrificing cosmetic results.

10.
Biomed Res Int ; 2020: 8926120, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32733961

RESUMO

Quantitative real-time PCR (qPCR) has become a widely used approach to analyze the expression level of selected genes. However, owing to variations in cell types and drug treatments, a suitable reference gene should be selected according to special experimental design. In this study, we investigated the expression level of ten candidate reference genes in hepatoma carcinoma cell (HepG2) and human hepatocyte cell line (L02) treated with ethanol (EtOH), hydrogen peroxide (H2O2), acetaminophen (APAP), and carbon tetrachloride (CCl4), respectively. To analyze raw cycle threshold values (Cp values) from qPCR run, three reference gene validation programs, including Bestkeeper, geNorm, and NormFinder, were used to evaluate the stability of ten candidate reference genes. The results showed that TATA-box binding protein (TBP) and tubulin beta 2a (TUBB2a) presented the highest stability for normalization under different treatments and were regarded as the most suitable reference genes of HepG2 and L02. In addition, this study not only identified the most stable reference genes of each treatment, but also suggested that ß-actin (ACTB), glyceraldehade-3-phosphate dehydrogenase (GAPDH), tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ), and beta-2 microglobulin (B2M) were the least stable reference genes in HepG2 and L02. This work was the first report to systematically explore the stability of reference genes in injured models of HepG2 and L02.


Assuntos
Regulação Neoplásica da Expressão Gênica , Fígado/lesões , Modelos Biológicos , Linhagem Celular Tumoral , Humanos , Padrões de Referência , Reprodutibilidade dos Testes , Software
11.
Biomed Res Int ; 2019: 6131879, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31223620

RESUMO

RAW264.7 is a macrophage strain derived from mice tumour and shows a significant ability in antigen uptake. Real-time quantitative PCR (RT-qPCR) is one of the most commonly used methods in gene studies and requires suitable reference genes to normalize and quantitate the expression of gene of interest with sensitivity and specificity. However, suitable reference genes in RAW264.7 cells have not yet been identified for accurate gene expression quantification. In the current study, we evaluated expression levels of ten candidate reference genes in RAW264.7 cells under different conditions. RT-qPCR results indicated significant differences in the expression levels among the ten reference genes. Statistical analyses were carried out using geNorm, NormFinder, and BestKeeper software to further investigate the stability of the reference genes. Integrating the results from the three analytical methods, cytochrome c-1 and hydroxymethylbilane synthase were found to be the most stable and therefore more suitable reference genes, while ribosomal protein L4 and cyclophilin A were the least stable. This study emphasises the importance of identifying and selecting the most stable reference genes for normalization and provides a basis for future gene expression studies using RAW264.7 cells.


Assuntos
Algoritmos , Perfilação da Expressão Gênica/normas , Expressão Gênica , Reação em Cadeia da Polimerase em Tempo Real/normas , Animais , Camundongos , Células RAW 264.7 , Padrões de Referência , Software
12.
Phytochemistry ; 164: 1-11, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31054374

RESUMO

Real time quantitative reverse transcription PCR (RT-qPCR) has been attracting more attention for its high sensitivity in gene expression analysis. Given the widely use of RT-qPCR in normalization, it is playing a pivotal role for seeking suitable reference genes in different species. In current work, 12 candidate reference genes including Actin 2 (ACT2), Cyclophilin 2 (CYP2), Glyceraldehyde-3-phosphate dehydrogenase C2 (GAPC2), Elongation factor 1-α (EF1-α), Nuclear cap binding protein 20 (NCBP20), Serine/threonine-protein phosphatase PP2A (PP2A), Polypyrimidine tract-binding protein 1 (PTBP1), SAND family protein (SNAD), TIP41-like protein (TIP41), Tubulin beta-6 (TUB6), Ubiquitin-conjugating enzyme 9 (UBC9) and Glyceraldehyde-3-phosphatedehydrogenase (GAPDH) were screened from the transcriptome datasets of M. charantia. Afterwards, GeNorm, NormFinder and BestKeeper algorithms were applied to assess the expression stability of these 12 genes under different abiotic stresses including drought, cold, high-salt, hormone, UV, oxidative and metal stress. The results indicated that 12 selected genes exhibited various stability across the samples under different external stress conditions, but TIP41, PTBP1 and PP2A presented high stability among all the reference genes. To validate the suitability of the identified reference genes, the results of hormone subset were compared with RNA sequencing (RNA-seq) data, and the relative abundance of Ascorbate peroxidase 1(APX1)was used to confirm the reliability of the results. This work assesses the stability of reference genes in M. charantia under different abiotic stress conditions, which will be beneficent for accurate normalization of target genes in M. charantia.


Assuntos
Momordica charantia/genética , Reação em Cadeia da Polimerase em Tempo Real , Perfilação da Expressão Gênica , Momordica charantia/crescimento & desenvolvimento
13.
Sci Rep ; 5: 18197, 2015 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-26657816

RESUMO

Despite comprising a small portion of the earth's surface, lakes are vitally important for global ecosystem cycling. However, lake systems worldwide are extremely fragile, and many are shrinking due to changing climate and anthropogenic activities. Here, we show that Poyang Lake, the largest freshwater lake in China, has experienced a dramatic and prolonged recession, which began in late September of 2003. We further demonstrate that abnormally low levels appear during October, 28 days ahead of the normal initiation of the dry season, which greatly imperiled the lake's wetland areas and function as an ecosystem for wintering waterbirds. An increase in the river-lake water level gradient induced by the Three Gorges Dam (TGD) altered the lake balance by inducing greater discharge into the Changjiang River, which is probably responsible for the current lake shrinkage. Occasional episodes of arid climate, as well as local sand mining, will aggravate the lake recession crisis. Although impacts of TGD on the Poyang Lake recession can be overruled by episodic extreme droughts, we argue that the average contributions of precipitation variation, human activities in the Poyang Lake catchment and TGD regulation to the Poyang Lake recession can be quantified as 39.1%, 4.6% and 56.3%, respectively.

14.
Sci Rep ; 4: 6600, 2014 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-25321660

RESUMO

While most large river-deltas in the world are facing the risk of subsidence and erosion in the Anthropocene, it is suspected that the Changjiang submerged delta (CSD) could be subjected to the impacts of the world's largest dam, the Three Gorges Dam (TGD). Here we firstly indicate that the CSD went through high accumulation (1958-1978); slight accumulation (1978-1997), slight erosion (1997-2002); and high accumulation (2002-2009), despite the 70% reduction of the sediment load from upstream since the operation of the TGD in 2003. Meanwhile, at the depocenter of the submerged delta, the accumulation maintained a high rate of 10 cm/yr during 1958-2009. This suggests on a longer term, the distal sediment source from the upstream had little effect on the CSD. Within this time frame the changes in the partition of sediment load among the branching channels of the Changjiang Estuary could likely control the shifting of the depocenter of the CSD on a decadal time scale. Episodic extreme floods and storm surges also increased the magnitude of deposition and erosion of the CSD on short-term scales. A re-evaluation of the impacts of TGD on the CSD is urgently needed.

15.
Sheng Wu Gong Cheng Xue Bao ; 28(7): 823-33, 2012 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-23167194

RESUMO

Studies on lycopene synthesis in Escherichia coli were not only able to gain the strains with high yield and less by-products, but also able to test functions of genes or gene clusters. In this article, the cDNA sequences of tomato LeGGPS2 and LePSY1 as well as the coding sequence of crtI from Erwinia uredovora, each of which was added a ribosome biding site, were controlled by T7 promoter and terminator alone or combined, and expressed in E. coli BL21 (DE3) to induce lycopene synthesis. The results show that only T7::crtI-LeGGPS2-LePSY1 expressed tri-cistronically could produce lycopene, and 2.124 mg/g dry cell weight oflycopene was obtained when fermented for 5 h at 30 degrees C after mixing 80 micromol/L IPTG at the later logarithmic phase while the seed broth of 1:50 (V/V) was inoculated into LB medium (pH 6.8) containing 3% sucrose and cultured for 8 h at 37 degrees C. The results confirmed the function of the prokaryonized LeGGPS2 and LePSY1 and their synergy with crtI, and also laid a foundation to establish an independent lycopene synthetic pathway in the tomato plastid.


Assuntos
Carotenoides/biossíntese , Erwinia/genética , Escherichia coli/genética , Proteínas de Plantas/genética , Proteínas de Bactérias/genética , Carotenoides/genética , Clonagem Molecular , Escherichia coli/metabolismo , Engenharia Genética , Licopeno , Proteínas de Plantas/biossíntese , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética
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