A novel series of first procaspase activating compound（PAC-1） analogues was designed, synthesized and evaluated for antitumor activity towards two cell lines[human promyelocytic leukemia cell line（HL60） and human ...A novel series of first procaspase activating compound（PAC-1） analogues was designed, synthesized and evaluated for antitumor activity towards two cell lines[human promyelocytic leukemia cell line（HL60） and human embryonic lung fibroblast cell line（HLF）] by the MTT[3-（4,5）-dimethylthiahiazo（-z-yl）-3,5-di-phenytetrazo- liumromide] method in vitro. The structures of all the compounds were confirmed by IH NMR, MS and elemental analysis. Among the compounds synthesized^（E）-2-[（3-{[4-（tert-buty~）benzy~]（methy~）amin~}pr^py~）（methy~）amin~]- N＇-[4-（diethylamino）-2-hydroxybenzylidene]acetohydrazide（compound 6n） exhibits a good anti-proliferative activity to the majority of tumor cells tested, and selectively cleaves cancer cells. Thus, compound 6n was identified as promising lead compound for further structural modification.展开更多
Abscisic acid(ABA)is a phytohormone that not only important for plant growth,but also mediating the stress response.The roles of ABA in plant immunity are especially multifaceted.Recently,the ABA functional analogues ...Abscisic acid(ABA)is a phytohormone that not only important for plant growth,but also mediating the stress response.The roles of ABA in plant immunity are especially multifaceted.Recently,the ABA functional analogues are of great significance to promote its application.Here,we reported an ABA functional analogue named 167A.167A inhibits plant growth and seeds germinating of Arabidopsis.Meanwhile,the 167A enhanced the plant immunity,which is opposite of ABA.We further investigated the PTI-response after 167A treatment,and the results show that the ROS burst,callose deposition accumulate with 167A treatment.Moreover,167A also influence the degree of stomal closed.RNA-seq assays show that the 167A down-regulated the ABA associated genes and upregulated the JA/SA/ET associated genes.Through genetic analysis,the 167A modulating the plant resistance through the PYR/PYL Receptors.Together,these results demonstrate that a novel ABA analogue 167A positive regulated plant immunity and has great potential for agricultural applications.展开更多
The Two-dimensional Quantitative Structure-activity Relationship （2D-QSAR） of a series of novel norcantharidin analogues, which exhibit hnhibitory activities of protein phosphatase 1 and 2A （PP1 and PP2A）, has bee...The Two-dimensional Quantitative Structure-activity Relationship （2D-QSAR） of a series of novel norcantharidin analogues, which exhibit hnhibitory activities of protein phosphatase 1 and 2A （PP1 and PP2A）, has been studied with a combined method of ab initio （I/F）, molecular mechanics （MM＋） and statistics. The established 2D-QSAR model （Eq. 1） for PP1 shows a reasonable regressive performance （R2= 0.749）, and the hydrophobic property of this molecule plays a decisive role in determining the inhibitory activity of PP1. In addition, the established 2D-QSAR model （Eq. 2） for PP2A also shows an acceptable regressive performance （R2= 0.701）, and the dipole moment of the molecule determines the inhibitory activity of PP2A.展开更多
The Quantitative Structure-Activity Relationship （QSAR） of a series of novel calanolide analogues,which exhibit inhibitory activities of HIV-1,has been studied with a combined method of ab initio （HF）,molecular me...The Quantitative Structure-Activity Relationship （QSAR） of a series of novel calanolide analogues,which exhibit inhibitory activities of HIV-1,has been studied with a combined method of ab initio （HF）,molecular mechanics （MM＋） and statistics. The established QSAR model （Eq. 1） shows a reasonable regressive performance （R2 = 0.885）. Both the surface area of the substituted group attached on C10,SR3,and the distance between atoms O13 and X14 （O,N,S）,L,of the calanolide analogues play important roles in determining the inhibitory activity of HIV-1.展开更多
BACKGROUND Fixed ratio combinations(FRCs)of analogue basal insulin and glucagon-like peptide-1 receptor agonists are a newer addition to the therapeutic armamentarium for the management of type 2 diabetes mellitus.The...BACKGROUND Fixed ratio combinations(FRCs)of analogue basal insulin and glucagon-like peptide-1 receptor agonists are a newer addition to the therapeutic armamentarium for the management of type 2 diabetes mellitus.They reduce treatment complexity by combining two injectables in a single daily injectable,thus potentially improving adherence and persistence.Clinicians wanting to use FRCs would need to choose between members of the class.AIM To describe and contrast the glycated haemoglobin reduction of two FRCs of analogue basal insulin and glucagon like peptide-1 receptor agonist in adults with type 2 diabetes mellitus.METHODS The following Population,Intervention,Comparison,Outcome question was used for the primary analysis:Among adult patients with type 2 diabetes mellitus[P],what is the effect of iGlarLixi[I]compared to IDegLira[C]for bringing about glycaemic control(as measured by reduction in glycosylated haemoglobin)[O]?The Prisma Statement was used as a guideline for framing this systematic review.We searched PubMed,EMBASE and Cochrane library databases and Clinicaltrials.gov using various keywords and medical search headings related to type 2 diabetes mellitus,iGlarlixi,IDegLira and glycated haemoglobin A1c.RESULTS All 14 studies identified by the systematic search met the primary efficacy endpoint of reduction in glycated haemoglobin.There were no head-to-head studies between the FRCs of iGlarlixi and IDegLira,and we therefore did an indirect comparison based on a common comparator of insulin glargine U100.Both iGlarLixi and IDegLira effectively reduce glycated haemoglobin when compared to insulin glargine U100.However,using indirect comparisons,IDegLira had a greater haemoglobin A1c reducing ability(0.6%vs 0.3%).The indirect comparison is limited by the differences between the studies;the fasting blood glucose targets were slightly higher for iGlarLixi studies when compared to the IDegLira studies(4.0-5.0 mmol/L and 4.4-5.6 mmol/L),and the IDegLira study used a greater average dose of insulin glargine when compared to the iGlarLixi studies(66 U/d vs 40 U/d).CONCLUSION Both iGlarLixi and IDegLira effectively reduce glycated haemoglobin.Indirect comparisons,using insulin glargine as the common comparator,suggest that IDegLira reduces glycated haemoglobin to a greater extent than iGlarLixi.However,given the limitations of indirect comparisons,robust head to head studies and real-world data would better inform clinician choice and clinical practice guidelines.展开更多
AIM: To investigate the interaction of reconstituted rhodopsin, 9-cis-retinal-rhodopsin and 13-cis-retinal-rhodopsin with transducin, rhodopsin kinase and arrestin-1. METHODS: Rod outer segments(ROS) were isolated fro...AIM: To investigate the interaction of reconstituted rhodopsin, 9-cis-retinal-rhodopsin and 13-cis-retinal-rhodopsin with transducin, rhodopsin kinase and arrestin-1. METHODS: Rod outer segments(ROS) were isolated from bovine retinas. Following bleaching of ROS membranes with hydroxylamine, rhodopsin and rhodopsin analogues were generated with the different retinal isomers and the concentration of the reconstituted pigments was calculated from their UV/visible absorption spectra. Transducin and arrestin-1 were purified to homogeneity by column chromatography, and an enriched-fraction of rhodopsin kinase was obtainedby extracting freshly prepared ROS in the dark. The guanine nucleotide binding activity of transducin was determined by Millipore filtration using β,γ-imido-(3H)-guanosine 5'-triphosphate. Recognition of the reconstituted pigments by rhodopsin kinase was determined by autoradiography following incubation of ROS membranes containing the various regenerated pigments with partially purified rhodopsin kinase in the presence of(γ-32P) ATP. Binding of arrestin-1 to the various pigments in ROS membranes was determined by a sedimentation assay analyzed by sodium dodecyl sulphatepolyacrylamide gel electrophoresis. RESULTS: Reconstituted rhodopsin and rhodopsin analogues containing 9-cis-retinal and 13-cis-retinal rendered an absorption spectrum showing a maximum peak at 498 nm, 486 nm and about 467 nm, respectively, in the dark; which was shifted to 380 nm, 404 nm and about 425 nm, respectively, after illumination. The percentage of reconstitution of rhodopsin and the rhodopsin analogues containing 9-cis-retinal and 13-cis-retinal was estimated to be 88%, 81% and 24%, respectively. Although only residual activation of transducin was observed in the dark when reconstituted rhodopsin and 9-cis-retinal-rhodopsin was used, the rhodopsin analogue containing the 13-cis isomer of retinal was capable of activating transducin independently of light. Moreover, only a basal amount of the reconstituted rhodopsin and 9-cis-retinal-rhodopsin was phosphorylated by rhodopsin kinase in the dark, whereas the pigment containing the 13-cis-retinal was highly phosphorylated by rhodopsin kinase even in the dark. In addition, arrestin-1 was incubated with rhodopsin, 9-cis-retinal-rhodopsin or 13-cis-retinal-rhodopsin. Experiments were performed using both phosphorylated and non-phosphorylated regenerated pigments. Basal amounts of arrestin-1 interacted with rhodopsin, 9-cis-retinal-rhodopsin and 13-cis-retinal-rhodopsin under dark and light conditions. Residual arrestin-1 was also recognized by the phosphorylated rhodopsin and phosphorylated 9-cis-retinal-rhodopsin in the dark. However, arrestin-1 was recognized by phosphorylated 13-cis-retinal-rhodopsin in the dark. As expected, all reformed pigments were capable of activating transducin and being phosphorylated by rhodopsin kinase in a lightdependent manner. Additionally, all reconstituted photolyzed and phosphorylated pigments were capable of interacting with arrestin-1. CONCLUSION: In the dark, the rhodopsin analogue containing the 13-cis isomer of retinal appears to fold in a pseudo-active conformation that mimics the active photointermediate of rhodopsin.展开更多
Phosphoinositide-dependent protein kinase-1 (PDK1), the class of serine threonine kinase, is a master regulator of the AGC family of kinases. It is a main component of the PI3K pathway. As it is reported that this pat...Phosphoinositide-dependent protein kinase-1 (PDK1), the class of serine threonine kinase, is a master regulator of the AGC family of kinases. It is a main component of the PI3K pathway. As it is reported that this pathway is most commonly, and this pathway is the most commonly deregulated among many cancers. So designing a selective inhibitor of PDK1 may have the efficacy as an anticancer agent. Herein, we describe our work focused on the structure based on screening of 95% similar analogues of Myricetin deposited in PubChem database as earlier studies have been suggested that myricetin acts as an anti cancer agent. Further molecular docking as well as the in silico ADMET studies are incorporated on these compounds to evaluate the binding and pharmacokinetic properties of these compounds. Due to low oral bioavailability, clinical use of myricetin is limited. Therefore this study is an attempt towards screening of structurally similar better compounds as compare with myricetin which can act as better inhibitor against PDK-1.展开更多
substituted hexahydro-1H-1,4-diazepine analogues were synthesized starting from N,N?-dibenzyl-1,3-propylene diamine and methyl-2,3-dibromo propionate through nucleophilic substitution, reduction, chlorination and debe...substituted hexahydro-1H-1,4-diazepine analogues were synthesized starting from N,N?-dibenzyl-1,3-propylene diamine and methyl-2,3-dibromo propionate through nucleophilic substitution, reduction, chlorination and debenzylation.展开更多
5-Substituted hexahydro-1H-1,4-diazepine analogues were synthesized starting from N,N'-dibenzyl-1, 2-ethylenediamine and methyl 2, 4-dibromide butyrate through nucleophilic substitution, reduction, chlorination, d...5-Substituted hexahydro-1H-1,4-diazepine analogues were synthesized starting from N,N'-dibenzyl-1, 2-ethylenediamine and methyl 2, 4-dibromide butyrate through nucleophilic substitution, reduction, chlorination, debenzylation and amidation. Bioactivity tests showed that 9a had the highest agonist activity.展开更多
The Schizosaccharmyces pombe pac\|1 gene product is a kind of dsRNA dependent ribonuclease,which has potential to degrade the dsRNA viral genome, the replication form of ssRNA viral genome and viroid genome.Therefore,...The Schizosaccharmyces pombe pac\|1 gene product is a kind of dsRNA dependent ribonuclease,which has potential to degrade the dsRNA viral genome, the replication form of ssRNA viral genome and viroid genome.Therefore,to introduce the pac\|1 gene into plants conferring them resistance to viruses is a new method of establishing the anti\|virus transgenic plant.The pac\|1 gene from the S.pmobe genome DNA isolated from China was cloned by means of PCR amplification.The pac\|1 gene was inserted into the cloning vector pGEM\|7Zf(+) by using restriction endonuclease Kpn Ⅰ/Bam HI.Sequencing analysis shows that it is a complete gene with 1095 necleotides.Compared to the reported pac\|1 gene,its homology is significant,but with 5 nucleotides differences,leading to only one amino acid difference. Pac\|1 gene was inserted into the prodaryotic expression vector pET\|21(a) by using the restriction endonuclase Nde Ⅰ/Bam HI.It was induced by the IPTG in E.coli BL21 harbouring the recombinant vector pET\| pac\|1 .The pac\|1 gene product is analyzed by the SDS\|PAGE.The result shows the product of pac\|1 gene exists in the supernatant part as soluble form and in the precipitant part as inclusion bodies after the cells were lysed by ultrasonic wave.The supernatant was applied to detect the enzyme activity of pac\|1 gene product.We conclued that pac\|1 gene has the biological activity of degrading the CMV\|dsRNA.展开更多
基金Supported by the National Natural Science Foundation of China（Nos.20771030, 20671025）.
文摘A novel series of first procaspase activating compound（PAC-1） analogues was designed, synthesized and evaluated for antitumor activity towards two cell lines[human promyelocytic leukemia cell line（HL60） and human embryonic lung fibroblast cell line（HLF）] by the MTT[3-（4,5）-dimethylthiahiazo（-z-yl）-3,5-di-phenytetrazo- liumromide] method in vitro. The structures of all the compounds were confirmed by IH NMR, MS and elemental analysis. Among the compounds synthesized^（E）-2-[（3-{[4-（tert-buty~）benzy~]（methy~）amin~}pr^py~）（methy~）amin~]- N＇-[4-（diethylamino）-2-hydroxybenzylidene]acetohydrazide（compound 6n） exhibits a good anti-proliferative activity to the majority of tumor cells tested, and selectively cleaves cancer cells. Thus, compound 6n was identified as promising lead compound for further structural modification.
基金supported by the National Key Research and Development Program(2022YFD1402100,2022YFD1401500)National Natural Science Foundation(32072500,32272557)+1 种基金Major Basic Research Project of Natural Science Foundation of Shandong Province(ZR2022ZD23)Shandong Province Key Research and Development Plan(2022TZXD0025,2021TZXD007-04-4),Taishan Scholar Program of Shandong Province.
文摘Abscisic acid(ABA)is a phytohormone that not only important for plant growth,but also mediating the stress response.The roles of ABA in plant immunity are especially multifaceted.Recently,the ABA functional analogues are of great significance to promote its application.Here,we reported an ABA functional analogue named 167A.167A inhibits plant growth and seeds germinating of Arabidopsis.Meanwhile,the 167A enhanced the plant immunity,which is opposite of ABA.We further investigated the PTI-response after 167A treatment,and the results show that the ROS burst,callose deposition accumulate with 167A treatment.Moreover,167A also influence the degree of stomal closed.RNA-seq assays show that the 167A down-regulated the ABA associated genes and upregulated the JA/SA/ET associated genes.Through genetic analysis,the 167A modulating the plant resistance through the PYR/PYL Receptors.Together,these results demonstrate that a novel ABA analogue 167A positive regulated plant immunity and has great potential for agricultural applications.
基金supported by the China Scholarship Council (CSC [2006] No. 3085)
文摘The Two-dimensional Quantitative Structure-activity Relationship （2D-QSAR） of a series of novel norcantharidin analogues, which exhibit hnhibitory activities of protein phosphatase 1 and 2A （PP1 and PP2A）, has been studied with a combined method of ab initio （I/F）, molecular mechanics （MM＋） and statistics. The established 2D-QSAR model （Eq. 1） for PP1 shows a reasonable regressive performance （R2= 0.749）, and the hydrophobic property of this molecule plays a decisive role in determining the inhibitory activity of PP1. In addition, the established 2D-QSAR model （Eq. 2） for PP2A also shows an acceptable regressive performance （R2= 0.701）, and the dipole moment of the molecule determines the inhibitory activity of PP2A.
基金supported by the Scientific Research Foundation of Department of Education of Yunnan Province ( No. 09Y0181)
文摘The Quantitative Structure-Activity Relationship （QSAR） of a series of novel calanolide analogues,which exhibit inhibitory activities of HIV-1,has been studied with a combined method of ab initio （HF）,molecular mechanics （MM＋） and statistics. The established QSAR model （Eq. 1） shows a reasonable regressive performance （R2 = 0.885）. Both the surface area of the substituted group attached on C10,SR3,and the distance between atoms O13 and X14 （O,N,S）,L,of the calanolide analogues play important roles in determining the inhibitory activity of HIV-1.
文摘BACKGROUND Fixed ratio combinations(FRCs)of analogue basal insulin and glucagon-like peptide-1 receptor agonists are a newer addition to the therapeutic armamentarium for the management of type 2 diabetes mellitus.They reduce treatment complexity by combining two injectables in a single daily injectable,thus potentially improving adherence and persistence.Clinicians wanting to use FRCs would need to choose between members of the class.AIM To describe and contrast the glycated haemoglobin reduction of two FRCs of analogue basal insulin and glucagon like peptide-1 receptor agonist in adults with type 2 diabetes mellitus.METHODS The following Population,Intervention,Comparison,Outcome question was used for the primary analysis:Among adult patients with type 2 diabetes mellitus[P],what is the effect of iGlarLixi[I]compared to IDegLira[C]for bringing about glycaemic control(as measured by reduction in glycosylated haemoglobin)[O]?The Prisma Statement was used as a guideline for framing this systematic review.We searched PubMed,EMBASE and Cochrane library databases and Clinicaltrials.gov using various keywords and medical search headings related to type 2 diabetes mellitus,iGlarlixi,IDegLira and glycated haemoglobin A1c.RESULTS All 14 studies identified by the systematic search met the primary efficacy endpoint of reduction in glycated haemoglobin.There were no head-to-head studies between the FRCs of iGlarlixi and IDegLira,and we therefore did an indirect comparison based on a common comparator of insulin glargine U100.Both iGlarLixi and IDegLira effectively reduce glycated haemoglobin when compared to insulin glargine U100.However,using indirect comparisons,IDegLira had a greater haemoglobin A1c reducing ability(0.6%vs 0.3%).The indirect comparison is limited by the differences between the studies;the fasting blood glucose targets were slightly higher for iGlarLixi studies when compared to the IDegLira studies(4.0-5.0 mmol/L and 4.4-5.6 mmol/L),and the IDegLira study used a greater average dose of insulin glargine when compared to the iGlarLixi studies(66 U/d vs 40 U/d).CONCLUSION Both iGlarLixi and IDegLira effectively reduce glycated haemoglobin.Indirect comparisons,using insulin glargine as the common comparator,suggest that IDegLira reduces glycated haemoglobin to a greater extent than iGlarLixi.However,given the limitations of indirect comparisons,robust head to head studies and real-world data would better inform clinician choice and clinical practice guidelines.
基金Supported by Grants from FONACIT,Caracas,Venezuela,No.S1-2000000514 and No.LAB-2000001639and from Decanato de Investigación y Desarrollo,Universidad Simón Bolívar,Caracas,Venezuela,No.S1-IN-CB-001-09
文摘AIM: To investigate the interaction of reconstituted rhodopsin, 9-cis-retinal-rhodopsin and 13-cis-retinal-rhodopsin with transducin, rhodopsin kinase and arrestin-1. METHODS: Rod outer segments(ROS) were isolated from bovine retinas. Following bleaching of ROS membranes with hydroxylamine, rhodopsin and rhodopsin analogues were generated with the different retinal isomers and the concentration of the reconstituted pigments was calculated from their UV/visible absorption spectra. Transducin and arrestin-1 were purified to homogeneity by column chromatography, and an enriched-fraction of rhodopsin kinase was obtainedby extracting freshly prepared ROS in the dark. The guanine nucleotide binding activity of transducin was determined by Millipore filtration using β,γ-imido-(3H)-guanosine 5'-triphosphate. Recognition of the reconstituted pigments by rhodopsin kinase was determined by autoradiography following incubation of ROS membranes containing the various regenerated pigments with partially purified rhodopsin kinase in the presence of(γ-32P) ATP. Binding of arrestin-1 to the various pigments in ROS membranes was determined by a sedimentation assay analyzed by sodium dodecyl sulphatepolyacrylamide gel electrophoresis. RESULTS: Reconstituted rhodopsin and rhodopsin analogues containing 9-cis-retinal and 13-cis-retinal rendered an absorption spectrum showing a maximum peak at 498 nm, 486 nm and about 467 nm, respectively, in the dark; which was shifted to 380 nm, 404 nm and about 425 nm, respectively, after illumination. The percentage of reconstitution of rhodopsin and the rhodopsin analogues containing 9-cis-retinal and 13-cis-retinal was estimated to be 88%, 81% and 24%, respectively. Although only residual activation of transducin was observed in the dark when reconstituted rhodopsin and 9-cis-retinal-rhodopsin was used, the rhodopsin analogue containing the 13-cis isomer of retinal was capable of activating transducin independently of light. Moreover, only a basal amount of the reconstituted rhodopsin and 9-cis-retinal-rhodopsin was phosphorylated by rhodopsin kinase in the dark, whereas the pigment containing the 13-cis-retinal was highly phosphorylated by rhodopsin kinase even in the dark. In addition, arrestin-1 was incubated with rhodopsin, 9-cis-retinal-rhodopsin or 13-cis-retinal-rhodopsin. Experiments were performed using both phosphorylated and non-phosphorylated regenerated pigments. Basal amounts of arrestin-1 interacted with rhodopsin, 9-cis-retinal-rhodopsin and 13-cis-retinal-rhodopsin under dark and light conditions. Residual arrestin-1 was also recognized by the phosphorylated rhodopsin and phosphorylated 9-cis-retinal-rhodopsin in the dark. However, arrestin-1 was recognized by phosphorylated 13-cis-retinal-rhodopsin in the dark. As expected, all reformed pigments were capable of activating transducin and being phosphorylated by rhodopsin kinase in a lightdependent manner. Additionally, all reconstituted photolyzed and phosphorylated pigments were capable of interacting with arrestin-1. CONCLUSION: In the dark, the rhodopsin analogue containing the 13-cis isomer of retinal appears to fold in a pseudo-active conformation that mimics the active photointermediate of rhodopsin.
文摘Phosphoinositide-dependent protein kinase-1 (PDK1), the class of serine threonine kinase, is a master regulator of the AGC family of kinases. It is a main component of the PI3K pathway. As it is reported that this pathway is most commonly, and this pathway is the most commonly deregulated among many cancers. So designing a selective inhibitor of PDK1 may have the efficacy as an anticancer agent. Herein, we describe our work focused on the structure based on screening of 95% similar analogues of Myricetin deposited in PubChem database as earlier studies have been suggested that myricetin acts as an anti cancer agent. Further molecular docking as well as the in silico ADMET studies are incorporated on these compounds to evaluate the binding and pharmacokinetic properties of these compounds. Due to low oral bioavailability, clinical use of myricetin is limited. Therefore this study is an attempt towards screening of structurally similar better compounds as compare with myricetin which can act as better inhibitor against PDK-1.
基金the Shanghai Development Fund of Science and Technology.
文摘substituted hexahydro-1H-1,4-diazepine analogues were synthesized starting from N,N?-dibenzyl-1,3-propylene diamine and methyl-2,3-dibromo propionate through nucleophilic substitution, reduction, chlorination and debenzylation.
文摘5-Substituted hexahydro-1H-1,4-diazepine analogues were synthesized starting from N,N'-dibenzyl-1, 2-ethylenediamine and methyl 2, 4-dibromide butyrate through nucleophilic substitution, reduction, chlorination, debenzylation and amidation. Bioactivity tests showed that 9a had the highest agonist activity.
文摘The Schizosaccharmyces pombe pac\|1 gene product is a kind of dsRNA dependent ribonuclease,which has potential to degrade the dsRNA viral genome, the replication form of ssRNA viral genome and viroid genome.Therefore,to introduce the pac\|1 gene into plants conferring them resistance to viruses is a new method of establishing the anti\|virus transgenic plant.The pac\|1 gene from the S.pmobe genome DNA isolated from China was cloned by means of PCR amplification.The pac\|1 gene was inserted into the cloning vector pGEM\|7Zf(+) by using restriction endonuclease Kpn Ⅰ/Bam HI.Sequencing analysis shows that it is a complete gene with 1095 necleotides.Compared to the reported pac\|1 gene,its homology is significant,but with 5 nucleotides differences,leading to only one amino acid difference. Pac\|1 gene was inserted into the prodaryotic expression vector pET\|21(a) by using the restriction endonuclase Nde Ⅰ/Bam HI.It was induced by the IPTG in E.coli BL21 harbouring the recombinant vector pET\| pac\|1 .The pac\|1 gene product is analyzed by the SDS\|PAGE.The result shows the product of pac\|1 gene exists in the supernatant part as soluble form and in the precipitant part as inclusion bodies after the cells were lysed by ultrasonic wave.The supernatant was applied to detect the enzyme activity of pac\|1 gene product.We conclued that pac\|1 gene has the biological activity of degrading the CMV\|dsRNA.
