Research of torque teno virus (TTV) in serum and total blood of Brazilian non-human primates and in chicken plasma (Gallus gallus domesticus) by the PCR N22 region / Investigación de Torque teno virus (TTV) en suero y sangre de primates no humanos brasileños y en plasma de pollo (Gallus gallus domesticus) por PCR en la region N22

Torque teno virus (TTV) is a recently discovered DNA virus that was originally isolated from a Japanese patient (initials, TT) with post-transfusion hepatitis of unknown aetiology. TTV is an circular DNA virus classified recently together with related Torque teño minivirus, into a new genus called Anellovirus. Infection TTV has been detected in a range of non-human primates as well as domestic animals. The purpose of this study was to search TTV in the serum and total blood of Brazilian monkeys and in plasma of domestic chickens by seminested PCR of coding region (N22), followed by a genomic sequence and phylogenetic analysis. No serum sample was amplified. TTV DNA was detected in total blood from 3 (4 percent) out of 75 brown-capuchin (Cebus apella) and from 1 (25 percent) out of 4 golden-headed lion-tamarin (Leontopithecus chrysomelas). Phylogenetic analysis revealed that one sample showed similarity with one sequence of the cotton top tamarin (Saguinus oedipus) (So-TTV2) and with one of the douroucoulis (ão tes trivirgatus) (At-TTV3). Two samples showed similarity with a human Torque Teño Mini Virus (TLMV). The other sample clustered with one sequence of the chimpanzee (Pt-TTV6) and with the human TTV strain TA278. The plasma chicken samples tested were all negative. The amino acid sequences reported in this study are the first obtained in Brazil from total blood of non-human primates naturally infected by TTV.

Torque teno virus (TTV) es un virus de ADN recientemente descubierto que fue inicialmente aislado de un paciente japonés (iniciales TT) después de la transfusión de hepatitis de etiología desconocida. TTV es un virus de ADN circular recientemente clasificado junto con los torque teno minivirus, en un nuevo género llamado Anellovirus. La infección de TTV se ha detectado en una serie de primates no humanos, así como animales domésticos. El objetivo de este estudio fue buscar TTV en el suero y sangre total de monos de Brasil y en el plasma de pollos domésticos, por seminested PCR de la región de codificación (N22), seguido de una secuencia genómica y el análisis filogenético. Las muestras que no eran suero fueron amplificadas. TTV DNA se detectó en sangre total de 3 (4 por ciento) de un total de 75 capuchinos de cabeza dura (Cebus apella) y de 1 (25 por ciento) de un total de 4 tití- león de cabeza dorada (Leontopithecus chrysomelas). El análisis filogenético demostró que una muestra presentaba similitud con una secuencia de Saguinus Edipo (So-TTV2) y con una de Aotes trivirgatus (A-TTV3). Dos muestras mostraron similitud con un torque teno mini virus (TLMV) humano. La otra muestra agrupada con una secuencia de los chimpancés (PT-TTV6) y con el TTV humanos cepa TA278. El análisis de las muestras de plasma de pollo fueron negativas Las secuencias de aminoácidos que se reportan en este estudio son las primeras obtenidas en Brasil de sangre de primates no humanos infectados naturalmente por TTV.

Purification and N-terminal Amino Acid Sequencing of the ESM Protease Isolated from an Eggshell Mem-brane-degrading Bacteria / 微生物学通报

A strain producing eggshell membrane protease (ESM protease) was isolated from the soil and identified as Pseudomonas aeruginosa. The enzyme isolated from the fermentation liquid of this strain and purified by ammonium sulfate precipitation, quadratic anion-exchange chromatography exhibited eggshell membrane degrading activity of 304.5 U/mg. By SDS-PAGE, the protein molecular mass is 32 kD. The N-terminal amino acid sequence of this protease is: Ala, Glu, Ala, Gly, Gly, Val, Ala, Gly, Lys, Glu, Asp, Ala, Ala, Glu, Leu.

Expression,Purification and Antibody Preparation of Recombinant C8orf32 Protein / 中国生物工程杂志

C8orf32 is a gene which has not been functionally characterized,the mRNA level of this gene is significantly higher in breast cancer tissues than that in normal breast tissues.The amplified cDNA fragment was inserted into the pGEX-6P1 vector fused with the upstream GST gene.The expression vector was transformed into the E.coli BL21(DE3) strain and expression of GST-C8orf32 fusion protein was induced by IPTG..After removal of GST tag by site-specific protease,the C8orf32 protein fused with an eight amino acid peptide tag was obtained.The purity of recombinant C8orf32 protein was about 95%.The identity of the purified protein was confirmed by N-terminal sequencing and tandem mass spectrometry.The polyclonal antibody was prepared by immunizing the New Zealand white rabbits with C8orf32 protein.The polyclonal antibody was proved to recognize the C8orf32 protein correctly.The purified C8orf32 protein can be used for structural and functional studies and the polyclonal antibody can be used for tissue specific protein expression profiling.

Identification of a novel osteosarcoma associated antigen 5D3Ag / 中国免疫学杂志

Objective:To analysis the N-terminal amino acid sequence of 5D3Ag purified by mAb5D3 immunoaffinity column.Methods:SDS-PAGE and Western Blot were used to study the characteristics of 5D3Ag.From SDS-PAGE gel,5D3Ag was transferred to PVDF membrane and its N-terminal amino acid sequence was analysised with an auto amino acid sequencer.Results:SDS-PAGE of 5D3Ag showed a band on the gel with molecular weight of 43 kD.Western Blot study showed that mAb5D3 can recognize 5D3Ag.The N-terminal amino acid sequence was NQGSHGSEEQ.Conclusion:It is sure that 5D3Ag is unique to Osteosarcoma because of the purification method itself and the characteristics of mAb5D3.Compared with other reported Osteosarcoma associated antigen,5D3Ag is different from them in molecular weight and may be a novel Osteosarcoma associated antigen.With the help of N-teminal amino acid sequence,some primers may be designed to clone the whole length gene of 5D3Ag.The identification of 5D3Ag gene may be useful in understanding the biologic mechanics of Osteosarcoma and developing new diagnostic and therapeutic strategies.