目的为快速、高效、准确地检出羊口疮病毒(ORFV),并进一步明确皖北地区羊群ORFV流行毒株的遗传变异情况。方法本研究基于ORFV F1L基因建立SYBR Green I实时荧光定量PCR(qPCR)方法,对皖北地区羊场开展ORFV检测,并从阳性病料中PCR扩增ORF...目的为快速、高效、准确地检出羊口疮病毒(ORFV),并进一步明确皖北地区羊群ORFV流行毒株的遗传变异情况。方法本研究基于ORFV F1L基因建立SYBR Green I实时荧光定量PCR(qPCR)方法,对皖北地区羊场开展ORFV检测,并从阳性病料中PCR扩增ORFV的完整VIR基因,直接测序后进行遗传演化分析。结果所建立ORFV SYBR Green I qPCR方法的线性关系良好,R 2=0.994;扩增效率高,E%=95.62%;检测速度快,理论反应时间约15 min;灵敏度较高,检测下限为10 copies/μL,是普通PCR法的100倍;重复性良好,组内和组间变异系数分别为0.44%~1.14%和2.82%~3.16%。用该qPCR方法对采集自皖北地区的168份羊临床样本进行检测,其ORFV阳性率为37.5%(63/168)。PCR扩增阳性样本中ORFV的完整VIR基因并直接测序,共获得52条全长VIR基因序列,其中21条来源于山羊,31条来源于绵羊。52条VIR基因间核苷酸序列及氨基酸序列的一致性分别为95.8%~100.0%和93.5%~100.0%,与参考毒株VIR基因核苷酸序列及氨基酸序列的一致性分别为95.1%~100.0%和91.3%~100.0%。基于ORFV VIR基因的遗传进化树分析显示,本次检测的21株山羊源ORFV毒株与8株国内外山羊源参考毒株和1株人源hChi17.2017参考毒株位于同一大分支,而本次检测的31株绵羊源ORFV毒株与7株国内外绵羊源参考毒株、1株人源hMbu15参考毒株及1株OV-V疫苗毒株位于遗传距离较远的另一大分支。结论建立了用于检测ORFV的高效、快捷、特异、灵敏、稳定的SYBR Green I qPCR方法。从临床样本中检测到的不同宿主来源ORFV的VIR基因间存在较大遗传差异。该研究结果可为ORFV的临床监测、检测、诊断及防控提供参考和指导。展开更多
Two kinds of signal factors capable of inducing Agrobaorerium vir gene expression were purified and identified from leaf extracts of panicle-differentiating to flowering stage of rice (Oryza saliva L. cv. IR 72) detec...Two kinds of signal factors capable of inducing Agrobaorerium vir gene expression were purified and identified from leaf extracts of panicle-differentiating to flowering stage of rice (Oryza saliva L. cv. IR 72) detected by Agrobacterium vir(?) lacZ. fusion genes. The induction was similar to that observed with 5 μm actosyringone (AS). Based on the comprehensive analysis of the data by UV, IR, NMR, MS, HMQC and HMBC, the structures of these two signal factors are identified as 5, 7, 4'-trihydroxy-3', 5'-dimethoxy-flavone (named tricin) and 5, 4' -dihydroxy-3', 5' -dimethoxy-7- (β-D-glucosyloxy) -flavone, respectively. These results demonstrate that monocotyledonous plants do contain highly efficient vir gene inducing factors of Agrobacterium, and the reason why monocotyledonous plants are difficult to transform by Ayrobacterium is not due to absence of vir gene inducing factors, but due to the signal factors only produced in specific stage and tissue of monocotyledonous plants展开更多
Introduction: This paper reports the results of a post marketing clinical study that tested the antiviral properties of Gene-Eden-VIRTM. Specifically, the clinical study tested the effect of Gene-Eden-VIR on the sever...Introduction: This paper reports the results of a post marketing clinical study that tested the antiviral properties of Gene-Eden-VIRTM. Specifically, the clinical study tested the effect of Gene-Eden-VIR on the severity, duration, and frequency of symptoms reported by individuals infected with various viruses. The viruses included the Human Papillomavirus (HPV), Herpes Simplex Virus (HSV), Epstein Barr Virus (EBV), Human Cytomegalovirus (HCMV) and Hepatitis C Virus (HCV). The symptoms included abnormal Pap smear, low and high grade cervical dysplasia, warts, blisters, cold sores, hives, skin tabs, panic attacks, depression, kidney problems, sleeping problems, liver problems, fever, fatigue, sore throat, swollen lymph nodes, diarrhea, and weight loss. Treatment: A capsule of Gene-Eden-VIR includes five natural ingredients: 100 mg of quercetin, 150 mg of green tea extract, 50 mg of a cinnamon extract, 25 mg of a licorice extract, and 100 mcg of selenium. The dosage was 1, 2, 3, or 4 capsules per day. The duration of treatment was 2 to 54 weeks. Population: The study population consisted of 60 infected individuals, ages 20 to 66. Results: The participants reported no side effects after taking Gene-Eden-VIR. Seventy three percent of the individuals treated with Gene-Eden-VIR reported a decrease in their symptoms. Specifically, they reported a decrease in the severity (p = 0.006, n = 45), duration (p = 0.009, n = 34), and frequency of their symptoms (p Conclusions: This post marketing clinical study showed that Gene-Eden-VIR is a safe and effective antiviral treatment. Specifically, the clinical study showed that Gene-Eden-VIR is a safe and effective treatment against the Human Papillomavirus (HPV), Herpes Simplex Virus (HSV), Epstein Barr Virus (EBV), Human Cytomegalovirus (HCMV), and Hepatitis C Virus (HCV). Therefore, health care practitioners should recommend Gene-Eden-VIR as a safe and effective antiviral treatment against these viruses.展开更多
Objective: The Microcompetition with Foreign DNA theory, proposed by Hanan Polansky in 2003, describes how latent viruses can cause chronic conditions, including fatigue. The Gene-Eden-VIR formula was designed to targ...Objective: The Microcompetition with Foreign DNA theory, proposed by Hanan Polansky in 2003, describes how latent viruses can cause chronic conditions, including fatigue. The Gene-Eden-VIR formula was designed to target latent viruses. Therefore, the theory predicts that treatment with Gene-Eden-VIR will decrease fatigue in individuals infected with a latent virus. The objective of this study was to test this prediction. Framework: A post marketing clinical study that followed FDA guidelines. Treatment: Gene-Eden-VIR, a dietary supplement. A capsule of Gene-Eden-VIR includes 100 mg of quercetin, 150 mg of green tea extract, 50 mg of cinnamon extract, 25 mg of licorice extract, and 100 mcg of selenium. The treatment included 1, 2, 3, or 4 capsules per day, and lasted 2 to 54 weeks. Population: 100 individuals infected with a latent virus, including the Human Papillomavirus (HPV), Epstein Barr Virus (EBV), Herpes Simplex Virus (HSV), Human Cytomegalovirus (HCMV), and Hepatitis C Virus (HCV). Ages ranged from 20 to 66. All participants reported a feeling of fatigue at the start of the study. Specifically, 98, 90, and 79 participants reported a feeling of general, physical, and mental fatigue, respectively. Results: Following treatment with Gene-Eden-VIR, 73.47%, 62.22%, and 47.36% reported a decrease in their feeling of general, physical, and mental fatigue, respectively. The participants also reported a statistically significant decrease in every aspect of fatigue tested in the study. The results also showed a duration effect, that is, those treated for 2 months or more reported a larger decrease in their feeling of fatigue (general, p = 0.03, n = 65;physical and mental, p = 0.05, n = 70). The results showed no interviewer bias, and no selection bias. In addition, the results showed therapeutic consistency under varying manufacturing conditions. The participants reported no side effects after taking Gene-Eden-VIR. Conclusions: This post marketing clinical study showed that treatment with Gene-Eden-VIR safely decreased the feeling of general, physical, and mental fatigue in individuals infected with a latent virus. Since most individuals are infected with a latent virus, health care practitioners should recommend Gene-Eden-VIR as a first line treatment for fatigue. The results of this post marketing clinical study support the Microcompetition with Foreign DNA theory.展开更多
Four rice (Oryza satia L. cv. IR72) signal factors with the functions of inducing Agrobaterium tumefaciens (Smith et Townsend) Conn vir genes were constructed. They were 5, 7, 4’-trihydroxy-3’, 5’-dimethoxy-flavone...Four rice (Oryza satia L. cv. IR72) signal factors with the functions of inducing Agrobaterium tumefaciens (Smith et Townsend) Conn vir genes were constructed. They were 5, 7, 4’-trihydroxy-3’, 5’-dimethoxy-flavone (F1) and its structure analogues F2, F3 and F4. These four signal fartors were detected by Agrobacterium vir:: lzcZ fusion gene system as to compare their functions of inducing virA, virB, virC,virD, virE and virG genes. The structure characters and structure-activity relationship of these signal factors with the best inducing effects to Agrobacterium exogenous gene transformation system were defined which may provide some basic information for foreign gene transformation in rice.展开更多
Either bacterial attachment or cellulose fibrillar elaboration was hardly observedduring the cocultivation of the cultured suspension cells of Oryza sativa Indica with thestrain C58C1 Rif^r of Agrobacterium tumefacien...Either bacterial attachment or cellulose fibrillar elaboration was hardly observedduring the cocultivation of the cultured suspension cells of Oryza sativa Indica with thestrain C58C1 Rif^r of Agrobacterium tumefaciens that was not specially pretreated. Onthe other hand, quite a lot of Agrobacterium cells were found to adhere to the surface ofcultured rice cells and a number of cellulose fibrils were produced around the specifiedbacteria when phenolics-pretreated bacteria were cocultivated with rice suspension cellsin common culture media, especially in complex culture solutions. The complex culturesolution was the bacterium-free filtrate of hormone-containing MS medium which hadbeen utilized to incubate carrot cells and the newly wounded hypocotyl segments fromtomato and Agrobacterium cells. Detecting experiments demonstrated that both NPT Ⅱ andNOS genes, located on the T-DNA segment of chimaeric plasmid pGV3850 :: 1103neo, weretransferred and expressed in the cultured cells of O. sativa Indica in the optimum trans-formation condition of the present experiment. Integration of the foreign genes into riceDNA was confirmed by Southern blot hybridization.展开更多
农杆菌已广泛应用于单、双子叶植物的遗传转化。T-DNA的传递是农杆菌介导转化的分子基础。T-DNA的传递是一个复杂的过程,与Ti质粒毒性区基因(Vir)、农杆菌染色体上毒性相关基因(chv)有关,另外植物体内部分基因也参与T-DNA的传递。T-DNA...农杆菌已广泛应用于单、双子叶植物的遗传转化。T-DNA的传递是农杆菌介导转化的分子基础。T-DNA的传递是一个复杂的过程,与Ti质粒毒性区基因(Vir)、农杆菌染色体上毒性相关基因(chv)有关,另外植物体内部分基因也参与T-DNA的传递。T-DNA传递时在VirD1-VirD2蛋白的作用下形成T链,进一步形成T复合物,经农杆菌四型分泌系统(Type IV secretion system,T4SS)穿过细菌和植物细胞膜,运输到植物细胞胞质。进入植物细胞的T复合物在植物相关蛋白的作用下经核运输和T链的整合,最终整合到植物基因组。本研究从农杆菌对植物细胞的识别和附着、农杆菌对植物信号的感知和Vir基因的活化、T链的形成、T-复合物的运输、T链进入植物细胞后的核运输和T链的整合机制及相关基因作了详细综述,探讨了农杆菌转化机制研究中的问题,对今后农杆菌转化机制研究做了展望。展开更多
文摘目的为快速、高效、准确地检出羊口疮病毒(ORFV),并进一步明确皖北地区羊群ORFV流行毒株的遗传变异情况。方法本研究基于ORFV F1L基因建立SYBR Green I实时荧光定量PCR(qPCR)方法,对皖北地区羊场开展ORFV检测,并从阳性病料中PCR扩增ORFV的完整VIR基因,直接测序后进行遗传演化分析。结果所建立ORFV SYBR Green I qPCR方法的线性关系良好,R 2=0.994;扩增效率高,E%=95.62%;检测速度快,理论反应时间约15 min;灵敏度较高,检测下限为10 copies/μL,是普通PCR法的100倍;重复性良好,组内和组间变异系数分别为0.44%~1.14%和2.82%~3.16%。用该qPCR方法对采集自皖北地区的168份羊临床样本进行检测,其ORFV阳性率为37.5%(63/168)。PCR扩增阳性样本中ORFV的完整VIR基因并直接测序,共获得52条全长VIR基因序列,其中21条来源于山羊,31条来源于绵羊。52条VIR基因间核苷酸序列及氨基酸序列的一致性分别为95.8%~100.0%和93.5%~100.0%,与参考毒株VIR基因核苷酸序列及氨基酸序列的一致性分别为95.1%~100.0%和91.3%~100.0%。基于ORFV VIR基因的遗传进化树分析显示,本次检测的21株山羊源ORFV毒株与8株国内外山羊源参考毒株和1株人源hChi17.2017参考毒株位于同一大分支,而本次检测的31株绵羊源ORFV毒株与7株国内外绵羊源参考毒株、1株人源hMbu15参考毒株及1株OV-V疫苗毒株位于遗传距离较远的另一大分支。结论建立了用于检测ORFV的高效、快捷、特异、灵敏、稳定的SYBR Green I qPCR方法。从临床样本中检测到的不同宿主来源ORFV的VIR基因间存在较大遗传差异。该研究结果可为ORFV的临床监测、检测、诊断及防控提供参考和指导。
基金Project supported by the Reckefeller Foundation's International Program on Rice Biotechnology.
文摘Two kinds of signal factors capable of inducing Agrobaorerium vir gene expression were purified and identified from leaf extracts of panicle-differentiating to flowering stage of rice (Oryza saliva L. cv. IR 72) detected by Agrobacterium vir(?) lacZ. fusion genes. The induction was similar to that observed with 5 μm actosyringone (AS). Based on the comprehensive analysis of the data by UV, IR, NMR, MS, HMQC and HMBC, the structures of these two signal factors are identified as 5, 7, 4'-trihydroxy-3', 5'-dimethoxy-flavone (named tricin) and 5, 4' -dihydroxy-3', 5' -dimethoxy-7- (β-D-glucosyloxy) -flavone, respectively. These results demonstrate that monocotyledonous plants do contain highly efficient vir gene inducing factors of Agrobacterium, and the reason why monocotyledonous plants are difficult to transform by Ayrobacterium is not due to absence of vir gene inducing factors, but due to the signal factors only produced in specific stage and tissue of monocotyledonous plants
文摘Introduction: This paper reports the results of a post marketing clinical study that tested the antiviral properties of Gene-Eden-VIRTM. Specifically, the clinical study tested the effect of Gene-Eden-VIR on the severity, duration, and frequency of symptoms reported by individuals infected with various viruses. The viruses included the Human Papillomavirus (HPV), Herpes Simplex Virus (HSV), Epstein Barr Virus (EBV), Human Cytomegalovirus (HCMV) and Hepatitis C Virus (HCV). The symptoms included abnormal Pap smear, low and high grade cervical dysplasia, warts, blisters, cold sores, hives, skin tabs, panic attacks, depression, kidney problems, sleeping problems, liver problems, fever, fatigue, sore throat, swollen lymph nodes, diarrhea, and weight loss. Treatment: A capsule of Gene-Eden-VIR includes five natural ingredients: 100 mg of quercetin, 150 mg of green tea extract, 50 mg of a cinnamon extract, 25 mg of a licorice extract, and 100 mcg of selenium. The dosage was 1, 2, 3, or 4 capsules per day. The duration of treatment was 2 to 54 weeks. Population: The study population consisted of 60 infected individuals, ages 20 to 66. Results: The participants reported no side effects after taking Gene-Eden-VIR. Seventy three percent of the individuals treated with Gene-Eden-VIR reported a decrease in their symptoms. Specifically, they reported a decrease in the severity (p = 0.006, n = 45), duration (p = 0.009, n = 34), and frequency of their symptoms (p Conclusions: This post marketing clinical study showed that Gene-Eden-VIR is a safe and effective antiviral treatment. Specifically, the clinical study showed that Gene-Eden-VIR is a safe and effective treatment against the Human Papillomavirus (HPV), Herpes Simplex Virus (HSV), Epstein Barr Virus (EBV), Human Cytomegalovirus (HCMV), and Hepatitis C Virus (HCV). Therefore, health care practitioners should recommend Gene-Eden-VIR as a safe and effective antiviral treatment against these viruses.
文摘Objective: The Microcompetition with Foreign DNA theory, proposed by Hanan Polansky in 2003, describes how latent viruses can cause chronic conditions, including fatigue. The Gene-Eden-VIR formula was designed to target latent viruses. Therefore, the theory predicts that treatment with Gene-Eden-VIR will decrease fatigue in individuals infected with a latent virus. The objective of this study was to test this prediction. Framework: A post marketing clinical study that followed FDA guidelines. Treatment: Gene-Eden-VIR, a dietary supplement. A capsule of Gene-Eden-VIR includes 100 mg of quercetin, 150 mg of green tea extract, 50 mg of cinnamon extract, 25 mg of licorice extract, and 100 mcg of selenium. The treatment included 1, 2, 3, or 4 capsules per day, and lasted 2 to 54 weeks. Population: 100 individuals infected with a latent virus, including the Human Papillomavirus (HPV), Epstein Barr Virus (EBV), Herpes Simplex Virus (HSV), Human Cytomegalovirus (HCMV), and Hepatitis C Virus (HCV). Ages ranged from 20 to 66. All participants reported a feeling of fatigue at the start of the study. Specifically, 98, 90, and 79 participants reported a feeling of general, physical, and mental fatigue, respectively. Results: Following treatment with Gene-Eden-VIR, 73.47%, 62.22%, and 47.36% reported a decrease in their feeling of general, physical, and mental fatigue, respectively. The participants also reported a statistically significant decrease in every aspect of fatigue tested in the study. The results also showed a duration effect, that is, those treated for 2 months or more reported a larger decrease in their feeling of fatigue (general, p = 0.03, n = 65;physical and mental, p = 0.05, n = 70). The results showed no interviewer bias, and no selection bias. In addition, the results showed therapeutic consistency under varying manufacturing conditions. The participants reported no side effects after taking Gene-Eden-VIR. Conclusions: This post marketing clinical study showed that treatment with Gene-Eden-VIR safely decreased the feeling of general, physical, and mental fatigue in individuals infected with a latent virus. Since most individuals are infected with a latent virus, health care practitioners should recommend Gene-Eden-VIR as a first line treatment for fatigue. The results of this post marketing clinical study support the Microcompetition with Foreign DNA theory.
文摘Four rice (Oryza satia L. cv. IR72) signal factors with the functions of inducing Agrobaterium tumefaciens (Smith et Townsend) Conn vir genes were constructed. They were 5, 7, 4’-trihydroxy-3’, 5’-dimethoxy-flavone (F1) and its structure analogues F2, F3 and F4. These four signal fartors were detected by Agrobacterium vir:: lzcZ fusion gene system as to compare their functions of inducing virA, virB, virC,virD, virE and virG genes. The structure characters and structure-activity relationship of these signal factors with the best inducing effects to Agrobacterium exogenous gene transformation system were defined which may provide some basic information for foreign gene transformation in rice.
文摘Either bacterial attachment or cellulose fibrillar elaboration was hardly observedduring the cocultivation of the cultured suspension cells of Oryza sativa Indica with thestrain C58C1 Rif^r of Agrobacterium tumefaciens that was not specially pretreated. Onthe other hand, quite a lot of Agrobacterium cells were found to adhere to the surface ofcultured rice cells and a number of cellulose fibrils were produced around the specifiedbacteria when phenolics-pretreated bacteria were cocultivated with rice suspension cellsin common culture media, especially in complex culture solutions. The complex culturesolution was the bacterium-free filtrate of hormone-containing MS medium which hadbeen utilized to incubate carrot cells and the newly wounded hypocotyl segments fromtomato and Agrobacterium cells. Detecting experiments demonstrated that both NPT Ⅱ andNOS genes, located on the T-DNA segment of chimaeric plasmid pGV3850 :: 1103neo, weretransferred and expressed in the cultured cells of O. sativa Indica in the optimum trans-formation condition of the present experiment. Integration of the foreign genes into riceDNA was confirmed by Southern blot hybridization.
文摘农杆菌已广泛应用于单、双子叶植物的遗传转化。T-DNA的传递是农杆菌介导转化的分子基础。T-DNA的传递是一个复杂的过程,与Ti质粒毒性区基因(Vir)、农杆菌染色体上毒性相关基因(chv)有关,另外植物体内部分基因也参与T-DNA的传递。T-DNA传递时在VirD1-VirD2蛋白的作用下形成T链,进一步形成T复合物,经农杆菌四型分泌系统(Type IV secretion system,T4SS)穿过细菌和植物细胞膜,运输到植物细胞胞质。进入植物细胞的T复合物在植物相关蛋白的作用下经核运输和T链的整合,最终整合到植物基因组。本研究从农杆菌对植物细胞的识别和附着、农杆菌对植物信号的感知和Vir基因的活化、T链的形成、T-复合物的运输、T链进入植物细胞后的核运输和T链的整合机制及相关基因作了详细综述,探讨了农杆菌转化机制研究中的问题,对今后农杆菌转化机制研究做了展望。
