Cloning and expression of the vp39 gene ofBombyx mori nuclear polyhedrosis virus inE. coli

The nuclear capsid protein gene (vp39) ofBombyx mori nuclear polyhedrosis virus (BmNPV) was amplified successfully by PCR technique and inserted into pGEM 3zf(+). The 5′ and 3′ terminal area of the amplified vp39 gene were sequenced with silver-staining dideoxy method. Bmvp39 gene was sub-cloned into the expression vector pRSET-A, and transformed intoE. coli BL21. This gene was highly expressed by IPTG induction. SDS-PAGE analysis showed that the expressed protein is about 38 kd, and the expressed amount reached maxium in 4 h with IPTG induction. Supported by the National Natural science Foundation of China and the Doctoral Foundation of Edn carto Committee Liu Deli: born in 1954, Doctoral Candidate from Huazhong Normal University To whom correspondence should be addressed: (027-7882712-2938)     

Cloning and expression of the vp39 gene of Bombyx mori nuclear polyhedrosis virus in E. coli

The nuclear capsid protein gene (vp39) ofBombyx mori nuclear polyhedrosis virus (BmNPV) was amplified successfully by PCR technique and inserted into pGEM 3zf(+). The 5′ and 3′ terminal area of the amplified vp39 gene were sequenced with silver-staining dideoxy method. Bmvp39 gene was sub-cloned into the expression vector pRSET-A, and transformed intoE. coli BL21. This gene was highly expressed by IPTG induction. SDS-PAGE analysis showed that the expressed protein is about 38 kd, and the expressed amount reached maxium in 4 h with IPTG induction. Supported by the National Natural science Foundation of China and the Doctoral Foundation of Edn carto Committee Liu Deli: born in 1954, Doctoral Candidate from Huazhong Normal University To whom correspondence should be addressed: (027-7882712-2938)     

Construction and<Emphasis Type="Italic">in vitro</Emphasis> study of an E1B-defective adenovirus

An E1B-defective adenovirus named rl/Ad was constructed by homologous recombination.The construction,selection and propagation of recombinant virus was done in the human embryonic kidney 293 cells (HEK293).The in vitro study demonstrated that the recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as the human glioblastoma tumor cells (U251) and human bladder tumer cells (EJ) but not in the normal cells with functional p53 such as the human fibroblast cells (MRC-5).Also,based on the cytopathic effect (CPE),it was demonstrated that the U251 cells were more sensitive to the infection of rl/Ad than that of EJ cells under identical conditions.In this paper,it was found that rl/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus.This may help to determine the safety of using any E1B-defective adenoviruses in cancer gene therapy.     

Expression of green fluorescent protein gene with baculovirus vector in insect cells

The green fluorescence of bioluminescent jellyfishAequorea victoria is due to the presence of the green fluorescent protein (GFP). To examine whether the GFP gene can be applied as a reporter gene in insect cells, a baculovirus transfer vector containing the neomycin resistance gene (neo) was established. The GFP gene was subcloned into the vector downstream of the polyhedrin gene (ocu) promoter. In the presence of G418, the recombinant virus can be purified. Expression of the GFP gene in the recombinant virus should give rise to synthesis of the GFP with a molecular weight of 30×10³ dalton, and is observable by the strong green light irradiated by ultraviolet or blue light in viable intact insect cells. The GFP produced in insect cells has typical fluorescent spectra indistinguishable from those of the purified native GFP. The GFP gene as a good reporter gene can be applied to the baculovirus-insect cell expression system. Supported by the National Natural Science Foundation of China Hu Jianhong: born in July, 1972, Master graduate student     

High expression of a rice homeobox inE. coli

Homeotic genes share a characteristic DNA segment, the homeobox, which encodes a defined domain of the homeotic protein-homeodomain which seems to mediate the binding to specific DNA sequences, whereby the homeotic protein exerts a gene regulatory function. In this study, the homeodomain encoded by the OSIH-1 (designated fromOryza sativa-Indica homeobox-1) homeobox of rice was overproduced in a expression vector inE. coli, as a form of inclusion body and analyzed by Western blotting and crossed-immunoreaction. Crossed-immunoreaction studies among OSIH-1, Kn-1 and Quox-1 indicate all three are homologous at the protein level. The OSIH-1 homeodomain is then to identify the homeotic target genes. Foundation item: Supported by the National Natural Science Foundation of China (#39570370) Biography: CHENG Xing-guo (1973-), male, Graduate student.     

A tumor suppressor genefhit prokaryotic expression and identification

The recombinant expression vector pGEMD-fhit which contains full encoding region offhit gene was constructed. The recombinant was introduced into the BL21 (DE3) strain ofE. coli and induced by 1 mmol/L IPTG to express a 29×10³ polypeptide offhit fusion protein. And the 29×10³ protein was sensitive and specific in reaction with anti-fhit antibody in Western blot. Foundation item: Supported by the National Natural Science Foundation of China (39770373) Biography: SUN Yan (1975−), female, Master of science, Research direction: gene engineering     

Physical location of the ricePi-5(t), Glh andRTSV genes by ISH of BAC clones

Mapping of low or single-copy sequences on plant chromosomes has proven difficult because of very low frequency of signal detection. Rice BAC library is being used widely in rice genome research due to its distinctive advantages over other library systems. In this study, two biotin-labeled rice BAC clones closely linked to a rice blast resistance, green leafhopper resistance and tungro spherical virus resistance gene,Pi-5(t), Glh, RTSV, werein situ hybridized to rice chromosomes. They were located on the long arm and short arm of chromosome 4 with FL value of 40% and 100% respectively. The frequency of signal detection reached 46.8% and 59.2%. The signal location were consistent with the selective marker on rice saturated molecular map. The results demonstrated the advantages to locate BAC clones to chromosomes byin situ hybridization and will facilitate the rice low or single-copy gene location by using the BAC library. Supported by the National Natural Science Foundation of China and the Doctorate Vesting Point Foundation of the Education Department of the People's Republic of China Yan Huimin: born in 1964, Lecturer     

Evaluation of some Fourier integrals inN-dimensional space for the theory of dislocations in quasicrystals

A method to evaluate some Fourier integrals is extended from two-dimensional (2-D) and three dimensional (3-D) spaces ton-dimensional (n-D) space, which are often used in the elasticity theory of dislocations in quasicrystals. Some key formulae have been given. Foundation item: Supported by the Natural Science Foundation of Hubei (9920p307) Biography: Yao Duan-zheng (1946-), female, Professor, Research interests: mathematical physics and nonlinear optics.     

Spatiotemporal characteristics of the energy radiation sources of the three great earthquakes near Sumatra Island in September 2007

Three strong earthquakes with magnitudes of Mw 8.4, Mw 7.9 and Mw 7.0 occurred in the sea west of Sumatra Island on September 12 and 13, 2007. We relocated the epicenters and focal depths of the three events by means of the reversal-time imaging technique using broadband digital seismic data from worldwide stations ranging from 30° to 90°, imaged the spatiotemporal variation of the energy radiation sources by means of the nonplane wave array technique using the broadband digital seismic data from a generalized array made up of 33 stations of the Capital Region Digital Seismograph Network （CRDSN）, and obtained the rupture duration times, extents and rupture velocities. Also, we discussed the correlations between the locations of the energy radiation sources of the three events.     

Study for interrelationship between human herpes simplex virus type 2 and leukemia

Two different strains of human herpes simplex virus type 2 (HHSV-2) and /or murine leukemia virus (MLV) were inoculated into newborn mice. In mice of mixed infection with MLV plus HHSV-2(333) or MLV plus HHSV-2(Wu)incidences of leukemia were respectively 79.16% and 75.00%,and in those inoculated with MLV alone were 41.02% and 44.74% in the experiment for 3 months. The differences of incidences between the two conditions were highly significant (p<0.01). On the contrary, no leukemia was detected in mice that were inoculated with HHSV-2 (333) or HHSV-2 (Wu) alone and in controls. Leukemia developing time of mice infected with two viruses was earlier than with MLV alone. It is obvious that the induction of murine leukemia resulted from infection of MLV and was independent of HHSV-2 (333) and HHSV-2(Wu). Both risen incidences and shortened latencies of murine leukemia were in close relationship with infection of HHSV-2(333) and HHSV-2(Wu). Foundation item: Supported by the National Natural Science Foundation of China (No. 59678017) Biography: Liu Jian-jun (1954-), male, associate research fellow, research direction: experimental oncology.     

The expression of vasa gene during zebrafish ( Danio rerio ) oogenesis

vasa gene expression pattern during oogenesis of zebrafish was examined usingin situ hybridization and fluorescent quantitative RT-PCR. During zebrafish oogensis,vasa mRNA is expressed strongly and uniformly distributed in the cytoplasm in stage II oocytes, followed by a distribution among vacuome in stage III. Later in stage IV and V,vasa mRNA is enriched at the cortex and finally localized at the cortex. The fluorescent quantitative RT-PCR shows that the quantity ofvasa mRNA decreases from stage II to stage III, but remains relatively invariable from stage III to stage V. The observed differences invasa mRNA expression in the different stages of zebrafish oogenesis suggest thatvasa gene plays an important role during oogenesis. Foundation item: Supported by the National Natural Science Foundation of China (30370744, 30150005) Biography: XIANG Fang (1979-), male, Master candidate, research direction: molecular development of animals.     

Evolutionary mode of metallothioneins inferred from Cd-MT genes of Tetrahymena

FromTetrahymena thermophila (strain BF5), the coding region ofCd-MT gene was cloned and sequenced, and identified as MTTI isoform. A serial duplicate structure is discovered in its amino acid sequence, which separates the coding region into three parts (Part 1: 7–61: Part 2: 64–118; Part 3: 122–162). The alignments among them and comparison with the corresponding parts of MT1 isoform suggest that MT1 and MTT1 isoforms both come from the same ancient gene that is homologous to Part 1, and Cd-MTs ofTetrahymena are aroused by such ancient gene duplication. The prediction of secondary structures and the analysis of the disulfide-bonding state of cysteine show that there are a lot of differences between MT1 and MTT1 isoforms, which maybe relate to their function mechanism. Foundation item: Supported by KSCX of Chinese Academy of Sciences Grant (SW-102) and the Allocation from the Earmaarked Grant for Hong Kong. Biography: WAN Mingliang (1980-). male, Ph. D. candidate, research direction: molecular ecotoxicology of protozoan.     

Baculovirus-mediated expression of a Chinese scorpion neurotoxin improves insecticidal efficacy

An Buthus martensii Karsch Insect Toxin （BmK IT ） gene was inserted into the genome of Autographa californica multicapsid nucleopolyhedrovirus （AcMNPV） to construct a recombinant baculovirus, AcMNPV-BmK IT. The expression of BmK IT was confirmed using RT-PCR, dot blot and SDS-PAGE analysis. Dose-lethal time responses to Spodoptera exigua larvae were compared between wild-type baculovirus AcMNPV and recombinant virus AcMNPV-BmK IT. At the concentration of 1 × 10^7 PIBs/mL, the median lethal time of recombinant baculovirus （LT50 = 73.6 h） on third instar S. exigua larvae showed an improvement of 13.2% over the efficacy of wild type virus （LT50 = 84.8 h） during a 192 h infection.     

Cloning and Characterization of the fecC Gene Necessary for Optimal Growth under Iron-Deficiency Conditions in the Cyanobacterium Anabaena sp. PCC 7120

The fecC gene encoding a putative iron (Ⅲ) dicitrate transporte rwas cloned from nitrogen-fixing cyanobacterium Anabaena sp. PCC 7120, and inactivated. The mutant grows normally in medium with NO3^- , NH1^- or without combined nitrogen. But in iron-deficient medium, the mutant grows slowly. Photosynthetic properties were compared between the mutant and the wildtype strain, the content of photosynthetic pigments in the mutant is lower than that of the wild-type. The results of RT-PCR experiments show that the fecC gene is expressed under iron-deficient conditions, but is not expressed under iron-replete conditions. These results revealed that fecC gene product is required for optimal growth under iron-deficient conditions in Anabaena sp. PCC 7120.     

NMR Revealed Activated Alumina-Water Interaction

Three different spin-lattice relaxation times ( T1 ) of water were obtained in activated alumina-water slurry system, which indicate that there exist three states of water: bound water, pore water and bulk water. The chemical shift (δ11) decreases as the amount of water added to the system increases due to the differences in contribution of these three states of water in the samples. The δ11 value for adsorbed water decreases nearly linearly and T1 increases with elevating temperature, which result from the decrease in the content of bound water by the increase in thermal motion.     

Search for new antiphytovirucides

A convenient method to synthesize substituted 2-(2′-hydroxyphenyl) benzimidazoles is reported. Six title compounds have been synthesized by the reaction of salicylic acid and 4-substituted0-phenylenediamine in the presence pyridine-POCl₃. Three compounds were tested as plant virucide against tobacco mosaic virus and they exhibited some activities. Supported by the National Natural Science Foundation of China, and National Laboratory of Elemento Organic Chemistry of Naikai University Huang Xiaoling: born in 1938, Professer     

Physical location ofHelminthosporium carbonum susceptibility gene hm1 by FISH of a RFLP marker umc119 in Maize

A fluorescencein situ hybridization (FISH) procedure was adopted to physically map a RFLP marker, umc119 near the centromere of the long arm of linkage group1 in maize. The hm1 gene (Helminthosporium carbonum susceptibility gene) was linked closely with the marker umc119. RFLP markers are very good landmarks for mapping genes. Therefore, we also determined the position of the gene hm1 on the chromosome based on the physical location of umc119. The disease induced by infection ofHelminthosporium carbonum is one of the serious maize diseases and it distributes in many countries including China. Hybridization sites were showed on 1 L (long arm of chromosome1) and 5 L. The percentage distance from centromere to the hybridization site was 22.86 on 1 L and 58.23 on 5 L the detection rate was about 12% for mitotic cells. In interphase nuclei five hybridized sites were detected. It demonstrated that umc119 was multiplicated sequences. FISH has more advantages overin situ hybridization (ISH) detected by DAB for increasing the detection ratio and contrast between chromosomes and hybridization signals. The ability to detect the hybridization signal of a small low copy DNA sequence is a very important key towards wide application of FISH for plant genome mapping. Supported by the National Natural Science Foundation and Doctorate Vesting Point Foundation of the Education Committee of China Li Lijia: born in 1967. Ph. D.     

Evolution of the serum resistance-associated SRA gene in African trypanosomes

Serum resistance-associated (SRA) protein, a protein unique for Trypanosoma brucei rhodesiense, is responsible for resistance of this parasite to the lysis by normal human serum (NHS) and is a vital molecular marker to distinguish this species from other African trypanosomes. We cloned and sequenced the SRA basic copy (SRAbc) gene from T. b. rhodesiense and related species and found that this gene is confined to the subgenus Trypanozoon. The average 82% identity among the sequenced SRAbc genes indicates that they may have a common origin and are highly conserved. Since SRAbc coexists in the T. b. rhodesiense genome with SRA, we propose that SRAbc might be the ‘donor VSG’, which after duplication became inserted into the expression site by recombination. Under natural selection, SRAbc could reform into SRA following mosaic formation. Supported by National Natural Science Foundation of China (Grant Nos. 30570245, 30670275), Changjiang Scholars and Innovative Research Team in University (Grant No. DPCKSCU/IRT0447), International Foundation for Science of Sweden (Grant No. B/4318-1), Grant Agency of the Czech Republic (Grant No. Z60220518) and Education Foundation of the Czech Republic (Grant No. 2B06129)     

On the approximation solvability of the abstract differential inclusion

The approximation solvability of the abstract differential inclusion du/dt∈f(t,u) is presented. The convergence of the approximation solution and the existence of the solution for abstract evolution multivalued problem are discussed. Foundation item: Supported by the National Natural Science Foundation of China (No. 19771062) Biography: Xia Jin-sheng (1975-), Ph.D candidate, research interest: the approximate solution of differential equation.