Isolation and Characterization of Phytoene Desaturase cDNA from Stigma of Crocus sativus

Phytoene desatumse (PDS) has recently been identified as an important enzyme in carotenoid biosynthesis pathway. A cDNA clone encoding phytoene desaturase gene is isolated from stigma of saffron (Crocus sativus L. ) using RT-PCR technique. Sequence analysis shows 85% similarity to Narcissus pseudonarcissus, 79% to Zea mays, 78% to Arabidopsis thaliana, 77% to Lycopersicon esculentum. A new full-length cDNA is obtained by 5 ‘-RACE and 3‘ -RACE techniques. The cDNA is 2149bp long with an open reading frame of 1697bp, which encodes a polypeptide of 565 amino acids. Southern analy-sis shows that the PDS gene is a single copy in saffron. Northern blot analysis shows higher expression level of PDS gene in stigma and anther than in leaves and stem.     

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     

Molecular cloning, expression and biochemical property analysis of AtKP1, a kinesin gene from Arabidopsis thaliana

Kinesins are common in a variety of eukaryotic cells with diverse functions. A cDNA encoding a member of the Kinesin-14B subfamily is obtained using 3＇-RACE technology and named AtKP1 （for Arabidopsis kinesin protein 1）. This cDNA has a maximum open reading frame of 3.3 kb encoding a polypeptide of 1087 aa. Protein domain analysis shows that AtKP1 contains the motor domain and the calponin homology domain in the central and amino-terminal regions, respectively. The carboxyl-terminal region with 202 aa residues is diverse from other known kinesins. Northern blot analysis shows that AtKP1 is widely expressed at a higher level in seedlings than in mature plants. 2808 bp of the AtKP1 promoter region is cloned and fused to GUS. GUS expression driven by the AtKP1 promoter region shows that AtKP1 is mainly expressed in vasculature of young organs and young leaf trichomes, indicating that AtKP1 may participate in the differentiation or development of Arabidopsis thaliana vascular bundles and trichomes. A truncated AtKP1 protein containing the putative motor domain is expressed in E. coil and affinity-purified. In vitro characterizations indicate that the polypeptide has nucleotide-dependent microtubule-binding ability and microtubule-stimulated ATPase activity.     

The microscopic approach of sdgIBM-1 and its application to nucleus<Superscript>154</Superscript>Gd

Based on the Dyson expansion theory, a microscopic approach of sdgIBM-1 is presented and applied to nucleus¹⁵⁴Gd in this paper. The energy spectra andE2 transition have been calculated. Good agreement is obtained in comparison with experimental results. Foundation item: Supported by the Foundation of Administration of Education of China Biography: Sang Jian ping (1959), male. Ph. D. Professor, research direction: nuclear physics     

Molecular cloning of a fulllength cDNA for ECBP21 from Angelica dahurica

ECBP21 is an extracellular calmodulin-binding protein which was first detected and purified from extracellular extracts of suspension-cultured cells of Angelica dahurica. The purified protein was electroblotted onto PVDF membrane and the amino acid sequences from 1 to 20 were determined. Using degenerate oligonucleotides of the sequence, a full-length cDNA coding for ECBP21 was isolated by a combination of RT-PCR and 5′-RACE cloning. The cDNA contains 947 nucleotides and codes for a precursor protein of 216 amino acids. The N-terminal 1–25 amino acid sequence is a predicted signal peptide and the other 26–216 amino acid sequence is a mature peptide. The 26–45 amino acid sequence shows identity with the N-terminal amino acid sequence of purified ECBP21 from Angelica dahurica. The fragment of encoding the mature protein was cloned into pET-28b(+) and transformed into E. coli BL21(DE3). A protein with relative molecular mass 21 ku was expressed in E. coli. Using a biotinylated-CaM gel overlay technique, the expression protein was tested for its ability to bind CaM. The results indicated that the expression protein is a Ca²⁺-dependent CaM-binding protein. Thus, these results further defined the cDNA clone for ECBP21. This work laid a foundation for elucidating biological functions of ECBP21 by using molecular biological means.     

SCFP,a novel fiber-specific promoter in cotton

To investigate the expression pattern of GhSCFP which was isolated from cotton fiber cDNA library, a 1006 bp upstream fragment of the gene was cloned by chromosome walking and fused to GUSand GFP respectively. Histochemical GUS and GFP fluorescence analysis revealed that the expression of the report genes driven by the promoter sequence was detectable only in outer layer cells during the seed development in the transgentic tobaccos. In transgenic cotton, strong GUS activity was observed in spherical protrusions on 0 dpa （days post anthesis） ovule surface, and in the 2-36 dpa fiber cells, while no GUS signals were detected in the root, leaves, stem, corolla, anther and stigma. Our data demonstrated that GhSCFP upstream sequence is a cotton fiber-specific promoter and this promoter will be useful in the molecular research on fiber cell development and in cotton fiber improvements by genetic modification.     

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.     

Cloning and Expression of BTB/POZ-Domain from Zebrafish gcl （Germ Cell-Less）, and Appraisement of Its Polyclonal-Antibody

In this study, a recombinant pET28c-gBTB/POZ was constructed by cloning the sequence of the BTB/POZ domain of the zebrafish gcl （germ cell-less） into the expression vector pET28c, and pET28c-gBTB/POZ was transformed into BL21（DE3） pLysS strain to express the fusion protein for the preparation of antibody. Polyclonal-antibody against the GCL-BTB/POZ domain was prepared by immunizing rabbit with the fusion protein, and the Western Blot and immuno-histochemical analysis were performed to detect the quantity of the polyclonal-antibody. The result indicates that the polyclonal-antibodies were of good quantity and specification. Further studies will be performed to demonstrate the function and expression pattern of the GCL protein during the development process of zebrafish with the polyclonal-antibody.     

Convergence in the<Emphasis Type="Italic">r</Emphasis>-th mean and the Marcinkiewicz type weak law of large numbers for weighted sums of<Emphasis Type="Italic">L</Emphasis><Subscript>q</Subscript>-mixingale arrays

L _r convergence and convergence in probability for weighted sums ofL _q-mixingale arrays have been discussed and the Marcinkiewicz type weak law of large numbers forL _q-mixingale arrays has been obtained. Foundation item: Supported by the National Natural Science Foundation of China (10071058) Biography: Gan Shi-xin (1939-), male, Professor, research direction; martingale theory, probability limiting theory and Banach space geometry theory.     

A Note on the Multifractal Decomposition of Directed Graph Self Similar Sets

We investigated the directed graph self similar sets under some weak overlapping condition. We get the multifractal decomposition formulas for these sets, i. e.,dim Ku^a DimKu^a =f(a), where fis the multifractal spectral function of the directed graph self similar measure. Especially, the results improve that of Edgar and Mauldin to the case which allows certain overlapping.     

HPV16E7-HSP70 Hybrid DNA Vaccine Induces E7-Specific Cytotoxic T Cells and Antitumor Immunity

0 IntroductionInbroicguhltat einxpge cDtaNtiAonvafcocri nper eivse ant ninegw aanpdp cruoraicnhg wdiits-heases . Due to the comparative weakness of cellulari mmunity that is purely induced by E7 DNA vac-cine,research on DNA vaccine is now focused onhowto enhance the i mmunogenicity of DNA vac-cine.It has been found in some research that thestrength of cellular i mmunity activity has things todo with the speed of antigen molecules’degrada-tion,i .e.the faster the degradation,the higher th…     

Study on the action of sodium selenite on the mitochondria metabolism of<Emphasis Type="Italic">Carassius auratus</Emphasis> hybrid carps liver by microcalorimetry

By using an LKB-2277 bioactivity monitor and ampoule method, the fundamental thermogenesis curves of the metabolic process of liver mitochondria fromCarassius auratus hybrid carps and the toxic effect of Na₂SeO₃ on it were studied at 28°C. From the thermogenesis curves, the thermokinetic equations were established under different condition. The kinetics show that Na₂SeO₃ has toxic action on the metabolism process ofCarassius auratus hybrid carps liver mitochondria. Foundation item: Supported by the National Science Foundation of China (29973030, 30170010 and 5020310), Chenguang Project of Wuhan and National Postdoctor Foundation of China Biography: Li Xi (1968-), female, Associate pofessor, research direction: biothermochemistry and biomaterials.     

Purification and Characterization of an Antibacterial Protein from the Cultured Mycelia of Cordyceps sinensis

0 IntroductionAlnattie mdicfrroobmiala p wriodteei nvsar iheatdy boefe linv ifnogun odr gaanndis ismos--Bacteria[1], fungi[2 ,3], plants[4]and ani mals[5].Those proteins displayed a wide spectrumof anti mi-crobial activity against different species of viruses ,bacteria andfungi .Over the past few years ,several anti microbialpeptides and proteins were foundinfungus ,such asAFP fromAspergillus giganteus[6], Anafp fromAspergillus niger[7], Zygocin fromthe yeastZy-gosaccharomyces bailii[8],an…     

Synthesis and Characterization of Polyamide-Containing 2, 6- （Substituded） pyridine Derivatives

0 IntroductionPayrriodminaetiics hoenteer oofc ythcleics m .os tC oambupnoduanndts a cnodn tbaeisntin kgno twhnepyridine ring are widely distributedin nature,principally asenzymes and alkaloids . Pyridine enzymes have been foundintissues of all plants and ani mals examined thus far ,andare derivedfromeither nicotinic acid or Vitamin B6[1].Pyr-idine derivatives are also the building block of many phar-maceuticals with a wide range of functionalities that includeantitubercular compounds ,antivi…     

Purification and inhibition fungal growth of chitinases from<Emphasis Type="Italic">Vibrio pacini</Emphasis>

Vibrio pacini synthesizes multiple chitinases, of which three have been purified in this study by ammonium sulphate fractionation, chitin affinity chromatography and gel chromatography. Molecular weights of the three chitinases, Chi1, Chi2 and Chi3 are 27×10³, 39×10³ and 46×10³ respectively, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The purified enzymes have optimal activity at pH 7–8, and retain 50% enzymatic activity pH 4–9. The activities of chitinases are inhibited by Pb²⁺, Fe³⁺ and Cu²⁺, and increased by Ca²⁺, Mg²⁺ and Mn²⁺. Chi3 is found to inhibit the growth of six species of fungi. Such characters of chitinase are different from those of any other chitinase that were reported before. Foundation item: Supported by the Key Technologies Research and Development Programme of the Tenth Five-Year Plan of the Nation Scientific and Technological Development (2001 BA708B04-07) Biography: HAN Bao-qin (1963-), female, Professor.     

Expression of TGF-β2 in LECs of Age-Related Nuclear, Cortex Cataract and the Relationship among TGF-β2, Proliferation,Apoptosis and Transdifferentiation

0 IntroductionAsgue-br-eclaaptseudl acrat acraatcatr ianctc l,ud aems ocnogrte xwh,incuhcl,e acro ratnexdcataract and nuclear cataract are more i mportant .Cortex cataract is different from nuclear cataract inoccurring,developing and clinic display,evenin his-tology. The pathogenesis of age-related cataract hasnot been perfectly described.But it has been verifiedthat thereis alayer oflens epithelial cells (LECs) un-der vertebrates’lens anterior capsule, which is i m-portant in maintaining t…     

Some Results about the Sample Path Properties of Markov Processes with Independent Self-Similar Components

This paper considers a special class of operator self-similar processes Markov processes {X（t）, t≥0} with independent self-similar components, that is, X （ t ） =（X^1（t）,…,X^d（t））, where {X^i（t）,t≥0}, i=1,2,…,d are d independent real valued self-similar Markov processes. By means of Brel-Cantelli lemma, we give two results about asymptotic property as t→∞ of sample paths for two special classes of Markov processes with independent self-similar components.     

Compression behavior and equation of state of Ni77P23 amorphous alloy

The compression behavior of Ni77P23 amorphous alloy is investigated at room temperature in a diamond-anvil cell instrument using insitu high pressure energy dispersive X-ray diffraction with a syn- chrotron radiation source. The equation of state is determined by fitting the experimental data accord- ing to Birch-Murnaghan equation: -ΔV/V0=0.08606P-3.2×10-4P2 5.7×10-6P3. It is found that the structure of Ni77P23 amorphous alloy is stable under pressures up to 30.5 GPa.     

On the Family of Thue Equation ｜x^3 ＋ mx^2y-（ m ＋ 3）xy^2 ＋ y^3｜= k

The family of cubic Thue equation which depend on two parameters ｜ x^3 ＋ mx^2 y-（m＋3） xy^2＋y^3｜=k is studied. Using rational approximation, we give a smaller upper bound of the solution of the equation, that is quite better than the present result. Moreover, we study two inequalities ｜ x^3 ＋ mx^2y-（m ＋ 3） xy^2＋y^3 ｜ =k≤2m＋3 and ｜x^3 ＋mx^2y- （m＋3）xy^2 ＋ y^3｜ = k≤ （2m＋3）^2 separately. Our result of upper bound make it easy to solve those inequalities by simple method of continuous fraction expansion.