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1994 Vol.09(4)



Universal Primer PCR Technology:A General Introduction of Applications

CHEN Huo-Sheng

1994, 9(4): 279

Ribozyme Used in Cleaving Nucleic Acid of Virus

CHEN Shi-You, JIN You-Xin

1994, 9(4): 283

Polymerase chain reaction for detection of different types of tiuman papillomavirus ill tile cervix

CAI Hong, TAO Kun, CHEN Wei-Li, ZHOU Yao-Xi, YANG Zhi-Jian

1994, 9(4): 291

Polymerase Chain Reactiun(PCR)was used to detect HPV 6,11,16,18 type specific DNA se-quences in 2l2 clinical cervical samples.The pusitivity uf HPV-DNA was 62.5%(25/40)in patientswith cervical cancer(squamous cell)and 57%(81/142)in chronic cervicitis,markedly higher than20%(6/30)in hedlthy control(P0.05).

Cloning and Expression of NS3 Gene Fragment of HCV and IdentlficatlOil of the Purified Products

LI Yue-Xi, TANG Jia-Qi, TAO Kai-Hua, JIAO Ren-Liang, LI Xian-Fu, PAN Xiu-Zhen

1994, 9(4): 297

The NS3 gene fragment of HCV was cloned from the serum of hepatitis C patients in Jiangshuprovince by RT-PCR.DNA sequencing shows the cloned NS3 cDNA contains 842bp,NcoI site and sal-I site were added respectively to the upetream and downstream primers.The cDNA was cut with NcoIand SalI,and then cloned into the NcoI and SalI sites in plasmid pBV221.The recombinant plasmidwas transformed into DH5a for gaining the E,coli expressing NS3 protein,Expressed NS3 protein(30kD)was about l7% of total proteins of the E.coli,and the purified NS3 protein by Sephacryl S-200 and IgG-Sepharose 4B chromatography(or DEAE Sepharose Fast Flow)has good antigenicity andspecificity.

Observation on the Antiviral Effect of HBIG .Acyclovir and Interferon Combining Treatment in Chronic Hepatitis B

HUANG Hua-Fang, XIONG Kai-Jun, ZENG Lian-Lan, HE Sheng-Song, YANG Ze-Chuan

1994, 9(4): 304

This paper reported the antiviral effect in 54 cases of chronic hepatits B,who were serum HBVreplication mark positive.It was divided into two groups:the treatment group and the control group in27 cases respoctively.In the treament group,acyclovir was used according to 25-20mg/kg/d in thefirst week,then became to 17-15mg/kg/d for 53 days intravenously;interferon was 1×106U im3/w for 4 weeks,then became to 1×106U im 2/w for 6 weeks;HIBIG was 400U im qod for l0 weeks.It was only treated with cummon protective hepdtic drugs in the control group.18 cases in thetreatment group and l9 cases in the control group had been fullowed-up half to two years;The resultshowed that the negative rate of HBeAg and HBV DNA in the recent and followed-up time reached40%,which was remarkable higher than the control group(P<0.05 ̄0.0l).4 and 2 cases with HBsAg became negative in the treatment group of different period,and none in the control group.Therecent and followed-up synthetical antiviral effect had observed that the whole negative

A Comparison of Four Wild Isolates of Heliothis Nuclear Polyhedrosis Virus

SUN Xiu-Lan, ZHANG Guang-Yu

1994, 9(4): 309

our Heliothis NPV isolates(two SNPV:HaS and HzS two MNPVs:HaMl and HaM2)were com-pared on the shape and structure,biological activity,structural polypeptide,restriction pattern andDNA homology.The LD50 values of the four isolates for late second to early third instar H.armigera larvae were 361,387,2633 and 3560 PIBs/g of diet;the LT50 valuse for 5×103 PIBs/g of diet were4.6,4.9,6.4 and 6.6 days respectively,Bioactivities of two SNPV isolates were higlier than that of two MNPV isolates.Assayed by SDS-PAGE,each polyhedrin of four isolates contained a singlepolypoptide.The structural polypeptides of HaM1 HaM2 comigated on SDS-gels,the strueturalpolypeptide maps of Has and HzS were similar,but there were little similarity between SNPV and MNPV structural polypeptides,Digested by BamHI、EcoRI、HindⅢ and XbaI,two MNPV genomes hadidentieal restriction patterns,and two SNPV genomes had similar fragmentation profiles,showing onlyslight difference in the number and size of several fragments.But SNPV and MNPV genomes had

Isolation of a M ild Strain of CMV with Satellite RNA and Cross Protection against Virulent Strains

ZHOU Xue-Ping, PU Zu-Qin, FANG Zhong-Da

1994, 9(4): 319

CMV isolate(CMVP 1)containing satellite RNA was obtained from pea(Pisum sativum)in Jiangsu province in 1990.Twenty-nine plant species representing nine families were inoculated mechanically with CMVP l and no symptoms or only mild symptoms were found.In Nicotina tabacum-Samsum,CMVP 1 caused Slight mosaic 8-16 days after inoculation and the infectivity redched the highestlevel at this time then the symptoms disappeared and the infectivity decreased.When satellite RNAwas eliminated by electrophoresis,CMVPl caused severe symptoms in tomato,so mild spoptoms of CMVPl in plant species were controlled by satellite RNA.In greenhouse crase protection tests,tomatoplants were first vaccinated with CMVP1 and cllallenge-inoculated with a virulent strain 5 to 30 dayslater,the disease incidence of vaccinated plant decreesed by 77.80%and the protective rate of the vaccination was higher than 90%,The average height and width of vaccinated plants were greater thanthose of healthy plants,so vaccination with CMVP1 had some favou

Study on Broad Bean Wilt Virus Isolated from Tomato

ZHOU Jun, YIN Yu-Qi, CUI Xing-Ming, LI Guo-Ying, LI Wei-Qi

1994, 9(4): 327

The isolate 87-35 was isolated from tomato in Xinjiang.After inoculating to tomato inducing mot-tle,the virus particles were isometric,and about 25nm in diameters,The virus infected 18 speciesplant-Chenopodium quinoa,C.amaranticolor,broad bean and tomato etc.It was transmitted by aphid(Myzus percicae)in nonpersistent manner.In the agar double-diffusion tests between the virus andbroad bean wilt virus antiserum,a clear band of precipitate was obseved,but it was not seen reactionbetween the virus and antisera of cowpea mosaic virus,and cucumber mosaic virus,Absorption spectrum of the virus was maximum in 258.5nm and minimum in 241.7nm.The virus particle containedtwo protein subunits,their electrophoresis molecular weights were about 45900 Dalton and 20400 Dal-ton reapectively,And they were composed of l8 kinds amino acids with 255 reeidues,Genome nucleicacid of the virus consisted of two components RNA,their molecular weights were about l320000 Dal-ton and 2340000 Daltun respectively.Accurding to the above resu

Molecular Cloning and Nucleotide Sequence of the Coat Protein Gene from Garlic Mosaic Virus

FAN Yong-Jian, WU Chu-Hua, LU Zhen-Xiao, XU Zheng

1994, 9(4): 333

They have isolated garlic mosaic virus(GarMV)from naturally infected garlic plants and synthesized various pattial cDNA fragments using the GarMV genomic RNA as template.The length of thecoat protein(CP)gene was estimated about 0.8 ̄0.9 kb based on the estimated molecular weight ofthe coat protein subunit by protein SDS-Polyacrylamide gel analysis,Several clones containing cDNAfragments larger than 2kb were searched for a 3'terminal cDNA fragment including the complete CPgene and noncoding region.One clone,pGM495 containing a 2.4kb fragment was identified to be related to G arMv genomic RNA by Northern blotting assay and to contain the 3'-terminal poly(A)tract by terminal sequencing analysis suggesting the presence of these regions.The cDNA fragment in pGM495 has been completely sequenced.It contains 2379bp and three u-nique restriction sites,EcoRI I Pst I and BamH I ,which are conistant with the primary reetriction en-donuclease mapping analysis.There are several in-frame stop cedons,the first stop codon TAA is

Studies of Cytological Effects Produced by Four viruses Assigned to the Furovirus Group

LI Yi, WEI Chun-Hong, PAN Nai, CHEN Zhang-Liang, TIAN Bo

1994, 9(4): 341

The cytological effects induced by beet necrotic yellow vein(BNYVV),beet soil-borne(BSBV),peanut clump African sourec(PCV-A),peanut clump Indian suurce(PCV-I)and soil-borne wheat mo-saic viruses(SBWMV)were studied.The four virusee,differed in the structure of membrane accumula-tions and/or the vesiculation of peroxisomes,Individual isolatea of a virus may differ in the cytologicaleffects induced.

Construction of Pseudorabies Virus Genomic Library and Analysis of Physical Mapping

WANG Ming-Shu, GUO Wan-Zhu, LOU Gao-Ming, BI En-Ge, XUAN Hua

1994, 9(4): 347

The Restriction endonuclease fragments except the ones from BamH I-1,2 were inserted into plasmid pBR322.The genomic library of Pseudurabies virus Min-A was also constructed.The Cloned frag-ment degested by BamH I were ldbeled with pliotobiotin,With molecular hybridization,the maincleavage site of Pseudorabies virus Min-A strain were determined.

Establishment of Hybridization Assay with Nucleic Acid Probe Detect IBDV and Comparison witIl Immunodiffusion Test

CHEN Shi-You, CHEN Pu-Yan, CA Bao-Xiang, JIN You-Xin, WANG De-Bao, ZHANG Ci-Jun

1994, 9(4): 351

The hybridization assay with digoxigenin(DIG)-labeled probe was developed for detection of in-fectious bursal disease virus(IBDV)and it's semitivity was compared with that of immunodiffusiontest in this experiment.IBDV STC-119 and STC-24 3 cDNA fragments used as probe were preparedfrom recombinant pUC plasmid with Hind Ⅲ and EcoRI and labeled with DIG.IBDV RNA was ex-tracted from IBDV purified from chicken bursa which were homogenized followed by extraction withchloroform,precipitation of virus with immune serum,digestion with proteinase K and recovery of IBDV RNA with ethanol precipitation.The results show that 0.lpg of IBDV RNA can be positively de-tected by this method and the mixed probe were able to hybridize with RNA from several IBDV isolatesand can be used for diagnosis of infectious bursal disease in the early stage of infection.The comparisonwith immunodiffusion test revealed that the furmer is l04 times more sensitive than the later,More-over,the specificity and the convenience of nonradioactive pro

Gone-protein—coding Assignments of Grass Carp Hemorrhage Virus Genome RNA Species

WANG Wei, CA Yi-Quan, FANG Qin, LIU Yuan-Wen, KE Li-Hua

1994, 9(4): 356

ll 11 segments of GCHV(grass.carp hemorrhage virus)873 dsRNA were isolated and individual-ly retrieved by SDS-PAGE and electric elution.Each viral genome segment in 90%DMSO denaturedby heating at 45℃ for l5 min were translated in the wheat germ tramlation system at 25℃ for 70min.The translation products were analysed in above system and l2 protein bands were displayed onthe X-ray film,RNA segments 1,2,3,4,5 and 10 coded individually for structural proteins of the in-ner virus shell,namely VP1,VP2,VP3,VP4,VP5 and VP10.RNA species coding for outer shell protenisVP6 and VP7 which are structural proteins were assigned to segments 6 and 7.Segments 8 and 9 codedindividually for VP52kD and VP 4l kD,and segment ll coded for VP29kD and VP 19.5kD.But weare not sure wether VP52kD,VP41kD,VP29kD and VP19.5kD are structural proteins or not.

Infection and M orphogenesis of Non-inclusion Body Baculovlrus from Pellaeus orientalis Kishinoye in Vivo

ZHANG Jian-Hong, CHEN Dai-Hua, XIAO Lian-Chun, ZHANG Li-Ren, GAO Xue-Xin, WU Zhi-Guang

1994, 9(4): 362

a lot of virogenic stroma,nucleocapsids,envelope and virion is detected in hepatopancreatic andmidgut epithelium cell nucleus of infected shrimp.Virion shapes short bar with blunt circle at twosides.The average size is 250-300×110nm.No inclusion body is deteeted in nucleus.There aresome two-layer-protein structures containing virion which we suggest be called occlusion-body,Mean-while,we think that this baculovirus without inclusion body could probably belong to the third sub-grope of insect baculoviride.

The Evaluation of the AVW C-test Effect Applied in the Life Science

DONG Chang-Yuan, CHEN Dong-E, WANG Sheng-Ji, YING Shi, CHEN Xiao

1994, 9(4): 367

AVWC-test is a new bio-statistical method which can transform the nonparameter datum into theparameter one to take the statistical analysis according to measurable datum.Some of the scienfific data were selected from the different research papers published in the academic magazine,analyzed statistically in the AVWC-test method as well as in other classical nonparametric statistic methods,And theself-control design were used to compare the statistical efficiency between the AVWC-test and the othertests.The result showes the AVWC-test is a excellent bio-statistical method because it is wide-ranging,accurate,sensitive and cunvenient so as to can better discover the scientific information than othermethods when you use it in biology field.

The Preparation of the Fluorescence Antibody of Ovine Progressive Pneumonia Virus and the Detection of the Cell Infected by the Virus


1994, 9(4): 375

The changes of sheep foetal lung cells infected by Ovine Progressive Pneumonia Virus(OPPV)were observed by.indirect immunofluorescence assay(IFA)and with the prepared fluorescence antibody of oppv.The result indicated that the fluorescence reaction initially appeared in 18 hours postinfection,The degree of the fluurescence reaction reached a high peak in 6 days postinfection and subsequently decreased.

Toxicity Test and Analysis of Genomic DNA of Spodoptera litura Nuclear Polyhedrosis Virus

WANG Xiao-Rong, LIU Ming-Fu, LIU Run-Zhong, LAN Ping-Zhang, ZHANG You-Qing

1994, 9(4): 378

The toxicity of a strain of Spodoptera tilura nuclear polyhedrosis virus(SINPV)was tested.The 4th instar larvae of Spodoptera titura were used to carryout the bioassays.LC5e=2.014×104PIBs/eachlarva.The genomic DNA was digested with three restriction endonucleases BamHI,EcoRI and PstI,and 17,22 and 12 fragments were produced,respectively,The total genome size is 113kb.