1994 Vol.09(1)



Cross-protection and its use on the control of plant virus disease

XIAO Huo-Gen, FAN Fu-Zhong

1994, 9(1): 1

Detection of I-IPV- 18 DNA with PCR

JIANG Gong, KE Li-Hua

1994, 9(1): 7

The DNA fragment(F)on the Upetream Regulatory Region(URR)of HPV-18 DNA was amplified with PCR.Two control experiments here confirmed the PCR specificity.The coneentration ofMg in PCR buffer affected mostly the specificity of PCR,the higher the coneentration of Mg,thelower the specificity,Moreover,nine out of seventeen(53%)DNA samples of cervical carcinoma tissues were detected with PcR for the presence of HVP-18 URR F sequences.It's implied that the re-sult may be an evidence of the relationship between HPV-18 and cervical carcinoma.

Study on expression of Epstein-Barr virus nuclear antigens in various infected states of cells

ZHAN De-Jin, CHEN Jian-Jing

1994, 9(1): 13

y microspectrophotometer quantitation(the expressing level of three kinds of EBNAs were detected with anti ̄EBNA antitheies purified from human serum by Western blotting method,The resultsshowed that EBNA-1 expressed greatly in latently infected Rajicell.When they were got into aborted-ly infected state treated with croton oil and n-Butyrate,the ex pression level of EBNA-1 decreased andthat of EBAN-2 increased,B_(95-8) cell had a similar variation,It suggeets that the activation of EB virusmight associated with the expression level of EBNAs.

Expression of HCV Core Protein in E.c0fi and Its Application

LI Yue-Xi, TANG Jia-Qi, SHI Jiang, ZHAO Hua-Zhong, JIAO Ren-Liang

1994, 9(1): 18

The cloned HCV core protein gene was inserted into the plasmid vector pJLA502 at EcoRI site,and the recombinant plasmid was transformed into E,coli DH_(5a),After inducing 5hrs at 42℃,SDS-PAGE analysis showed that the expressed core protein is twenty percent of total protein of E.coliDH5a;the purified core protein by Sephadex G200 gel filtration and Sepharose-4B-IgG affinity chro-matography has excellent antigenicity and specificity,and is good for detecting anti-HCV.

Expression HCV core protein in E. co//and its application

TANG Jia-Qi, LI Yue-Xi, LI Xian-Fu, PAN Xiu-Zhen, GUO Heng-Bin, ZHANG Yun

1994, 9(1): 25

My analysis and comparison of nucleotides sequences of HFRSV 76/118 and R_(22) strains,tlireepairs of primers were designed and synthesised,one pair of primer lying in the high homologous regionbetWeen 76/18 strain and R_(22) strain was used as common and outer primers;the other two pairs ofprimers were in the low homologous region,as the type-specific and inner primers.Using above primers and RT-PCR techneque,we measured five strains of HFRSV,76/118,A9,Chen,R_2,and R_(22).When using the outer primers,all of the five strains produced one DNA lane of 300bp;using the field-rat type inner primers all strains but R_(22) strain preduced one DNA Iane of 70bpand using the hom-rat type inner primers,only R_(22) strain produced one DNA lane of 70bp.Part of M fragment cDNA of 76/118 and R_(22) strains was used respectively as template,two type-specific biotinylated probed were synthesised by nest PCR techneque,the probes were used to hybridization with the RT-PCR preducts of the five strains,the results showed:RT-PCR te

Discussion Oil the Relationship between the Serum HBV Replication M arks and Hepatic Pathological Changes in the Chronic Hepatitis B of High RepBcafion Type

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

1994, 9(1): 31

The relationship between the serum HBV replication marks and hepatic pathological changes wasreported in 55 cases of chronic hepatitis B of high replication type.55 cases were HBsAg,HBeAg andanti-HBc positive,some accompanied with HBcAg,DNAP or HBV-DNA positive.All cases had madeliver biopsy and pathological test.The pathological report was:CAH 5 casea,CLH 9 cases and CPH 41cases.The first two states became Group A,which represented pathological change active,CPH became Group B,which represented inactive,Under the HBV replication state,the examined rate of thehepetic pathological change in the Group A was commonly higher than G roup B. Under the ALT abnor-mal state,the examined rate up to 80% of some hepatic patliological changes in Group A was alsohigher than Group B,It also indicated that piecemeal necrosis appeared in the 4 cases of Group A,andnone in Group B,Combinin8 the above result,it ind icated that there was close relationship among theHBV repl ication degree,the activity and extension of the hepotic

The Analysis on Esterascs from BsNPV-Infected Buzura suppreSSaria and Bs484 Ceils

XU Xu-Shi, ZHANG Chao-Yang, XIE Tian-En

1994, 9(1): 37

By the polyacrylamide gel electrophoresis(PAGE)and thin-layer scanning,the esterases of thefourth instar larvae of the Buzura suppressaria and Bs484 cells were investigated after infection with nuclear pelyhedrosis viruses,The amount of the zymograms for mid-gut esterases in diseased larvae wasfound to be less than that in the healthy ones during the course of the disease,The results showed thatthe changes in type and content of mid-gut esterases were investigated at 8 hours postinfection,and thecharlges became obvious as infective time went on,The changes in content of esterases for Bs484 cellswere determined at 3 hours postinfection with BsNPV,but the type of esterases had no changes.

Specific nucleic acids in host plant infected by Indian cassava mosaic geminivirus

HONG Yi-Guo, WANG Xiao-Feng, TIAN Bo

1994, 9(1): 43

J.Rovinson B.D.Harrison(Scottish Crop research Institute, Dundee,DD2 5DS,U.K.)Nucleic acids extracted from Indian cassava mosaic geminivirus infected plant leaves(Nicotiana ben thamiana)and further purified through 1.0% LMT agarose gel electrophoresis were treated with RNA ase,DNAase,Nuclease Sl,Exonuclease Ⅲ and restriction endonuclease EcoR I.Two kinds ofvirus-specific DNA existed in infected plants as shown by Southern and Dot blotting using virus ssDNAprobes synthesized by nick translation,These two DNAs were circu lar double stranded and singlestranded DNA(dsDNA and ssDNA)respectively.The dsDNA was in the form of open circular and covalently closed circular molecules(ee and ccc dsDNA).Both oc and ccc dsDNA were digested byEcoR I to produce lincar molecules of abeut 2.7 kb,The virus genome contained two components(DNA1 and DNA2)Furthermore,the circular viral ssDNA in plants was largely the miunus strandednot the plus stranded ssDNA reported from particles,This may be an important clue to the understandin

Detection of grapevine fanleaf virus by dot-blot hybridization with biotin-labelled GFV-cDNA probe

GU Hong-Cang, YAN Dun-Yu, LIU Huan-Ting, QIU Bing-Sheng, WANG Jin-Fang, TIAN Bo

1994, 9(1): 48

The 5′and 3'-primers of 21 bp of GFV coat protein gene were synthesized according to the GFV-RNA sequence,The cDNA of GFV ccat protein gene was syntheeized with reverse transcriptase using3′-primer and GFV-RNA extracted from the Durified virus as template,Then the ds-cDNA was amplified by PCR technique employing GFV-RNA_2:ss-cDNA as templates and 5',3′-primers and ligated-with pBluesclpt ks.The ds-cDNA were transformed into Ecoli XLI-Blue cells,g the cDNA extracted from the recombinantclones of E. coli cells.The biotin-labelled ss,ds-probes have been used for the detection of purified GFV-RNA,GFV-infeted C,quivoa and 18 grapevine leaves by nucleic acid dot-blot hybridization,The minimal amount ofpurified GFV-RNA_2 which gave the visible signal was 1.5pg/test dot and the maximum dilution ofGFV-infected C,quinoa extract which gave visible signal was 1:40960;11 samples with typical symptoms were,petive and some of them could give the signal when they were diluted 1:400 ̄1:800.3samples out of 7 symptomeless leaves

Using satellite RNA of cucumber mosaic virus (CMV) as a biological control agent to protect the wheat stubble pepper from disease in large area

WANG Zhi-Xue, DING Yu-Ying, QIN Bing-Yi, ZHANG Xiu-Hua, TIAN Bo, TIAN Wen-Hui

1994, 9(1): 54

The popper plants preinoculated with CMV satellite RNA biological control agent S52 were resiatant to infection ot CMV,The result of the field test in large area indicated that when pepper plantspreinoculated with S52 the protection effect was 59.5-71.0% compared with control plants,thefruit yield was increased by 26.1-32.3% and the output value was increased about 33.5-45.2%.While the result of the field test ln small area showed that pepper plants preinouclated with S52,showed a protection effect of 71.3-92.9% and 55.9-78.5% at 50 and 80 days postinoculation rospectively,the fruit yield and output value were 36.0-65.3% and 49.3-93,5% more than that ofthe control.The biological control agent also has some favourite effects on the plants such as stimutinggrowth,promoting early-maturation,and increasing resistance to fungal and bacterial diseases.

The Study on Two Virus Isolates of Beet in Xinjiang

CUI Xing-Meng, GONG Zu-Xun, YU Ning-Jiang

1994, 9(1): 60

Two virus isolates were found in beet in Xinjiang.After they are inoculated to beet the isolates induced yellow ring spot and line patterns along vein of leave.The rmgspot and line patterns eventuallyformed necrosis.The virus infected Chenopodium amaranticolor C.quinoa, Telragnia axpansa and Spinacia oleracea and induced local necrotic leeions.The virus infected Nicltiana tabacum,N. glolionosa,Pelunia hy brida,bean,cucumber and tomato but without showing symptoms.The virus particles have icosahedramorphology with an averege diameter of 28nm,In sap the virus thermal inactivation point(10min)is70℃.Infectivity is retained for 13 days at room temperatue about 20℃ and for 7 years in refrigerat ed dry leaves.The highly purified virus is easily obtained from C.amaranticolor by precipitation withPEG and differential centrifugation,Absorption spetrum of the virus is maximum in 260nm and minimum in 245nm.Protein subunits have M,W.27000.Cenome nucleic acid of the virus consists of three species with sizes of 3.08kb,1.28k

Ultrastructural Study of Molluscum Contagisum Virus(MCV)

LI De-Zhong, XIAO Tong-Hao, WU Xiao-Hua, WU Ning

1994, 9(1): 67

5 blocks of MC tissues from 15 patients were oteerved with TEM,MCV has 4 patterns,i.e,primary,middle'mature and degenerate phases,The primary viruses have teeny-granular- chain-likeviroptasm,and the middle ones have vironucleoid construction, condensed viroplasm and lateral bodies(LB).The mature McV is flat ellipsoid,the size is 400×300×180nm,and has no LB in this phase.The cores of mature MCV are of cubically biconcave ellipsuid(or elliptocyte-like)shapos,not dumhbell-like,and appear the concentric fingerprint-like structures in level section, Eventually,MCV becomes degenerate along with the death of the cells.

The Application of Rapid HEPES-plaques-forming Test(RHPFT)to Rabies Virus Titration

SHAO Yi-Bin, GU Qin, ZENG Rong-Fang

1994, 9(1): 70

According to the phenomenon that HEPES could enhance CPE in rabies virus infected cell and accelerate the plaques formation,the virulence of rabies virus(RFD strain)was detected with 3 differentmethods includig RHPFT,MC-plaques test and mollse i.c,test.The results showed that the sensibility of 3 detecting methods was similar but RHPFT was more rapid and simple than the latter two,Theresult of RHPFT methed could be obtained earlier about 6-7 days than MC-plaques test which everused as a routine assay in rabies virus titration.RIlPFT could also be used for rabies neutralizing anti-body assay.

Isolation and Identification of Paroceneria furva Nuclear Polyhedrosis Virus

LI Chong-Rong, PANG Hui-Yin, ZHOU Xian-Ming, CHEN Xin-Wen, XIE Tian-En

1994, 9(1): 74

The paper reports paroceneria furva single nuclear polyhedrosis virus(PfNPV)that was not described before.The size of mlyhe0m inclusion bodies(Pros)raiages from 1.0um to 1.35um.The virions are rod—shaped,approximately 371× 73nm.

Comparison of Tissue Culture and Hemagglutinalion between Four Animal Viruses

LI Tian-Xian, ZHAO Lin, LUO Yi-Shan, FENG Feng

1994, 9(1): 77

All of the Laopard Parvovirus(LPV), Enteritis Virus(MEV).Caine Parvovirus(CPV)8nd Feline Panleukopenia Virus(FPLV)can be bred on. l cel1.At the meantime,two different Hemagglutinations a” taken and comp lied. Through e) rimen ,we have found one HA method which simple·accuraleand stable.

Observation of Pathological and Viral Antigen of Epidemic-Hemorrhagic Fever Virus Infection in Suckling Mice

GUO Guang-Song, XIAO Hong, CHENG Li, WEN Li, YANG Zhan-Qiu

1994, 9(1): 81

Epidemic hemorrhagic fever virus origined from patients with epidemic hemorrhagic fever was inoculated into suckling mice,The results showed that the suckling mice died regularly,The viral antigenand pathological damage appeared in the brain,lung,liver and kidney,The characterization of patho-logical changes were congestion, hemorrhage and degeneration of blood vessels.The viral perticles canbe found in hippocampus region,The viral antigen can be detected in cytoplasm.The resultS suggestthat suckling mice were susceptible for human original epidemic hemorrhagic fever virus.