the ability to biosynthesis of protein A from some Staphylococcus aureus strains isolated from the patients of Bach Mai Hospital was studied in the years 2001-2002. The results showed that there were 3 strains (25, 28 and 55) have the ability to synthesize protein A in the highest levels among 8 strains investigated. Using ELISA technique with protein A served as an antigen from Staphylococcus strain 25 to analyse IgG from sera of 32 healthy subjects has determined that the average IgG level from human serum is 14.37mg/ml. This result was supported by the same technique using commercial foreign bio-kit. It is proven that protein A preparation can be used as an antigen in bio-kit capturing specifically IgG from human serum in clinical diagnosis and in immuno-pathological research using ELISA technique
Staphylococcal Protein A ; biosynthesis ; Enzyme-Linked Immunosorbent Assay

L-homophenylalanine (L-HPA) is an important non-natural amino acid that has been used as a key intermediate for the synthesis of Puli drugs for the treatment of hypertension. At present, L-HPA is synthesized using chemical methods, which has the disadvantages of expensive raw materials, tedious steps and serious pollution. Therefore, researchers have conducted in-depth research on the enzymatic production of L-HPA. This review summarizes the research progress on the enzymatic synthesis of L-HPA, including the dehydrogenase process, the transaminase process, the hydantoinase process, and the decarboxylase process, with the hope to facilitate the industrial production of L-HPA.
Amino Acids ; Environmental Pollution ; Industry ; Protein Biosynthesis

Translationally controlled tumor proteins (TCTP) and SNF1- related protein kinase (SnRK1) are conserved and widely present in eukaryotic cells. TCTP regulates cell division, plant growth and development, and mediates plant resistance against pathogen infection. SnRK1 participates in a range of physiological processes including sugar metabolism and resistance to abiotic and biotic stresses. Previous work in our laboratory demonstrated that wheat TCTP can respond to Puccinia triticina infection and induce host defense responses. In order to further investigate the mechanism of TaTCTP in wheat resistance to Puccinia triticina infection, we used TAP (tandem affinity purification) and mass spectrometry to screen the potential interactants of TaTCTP. A SNF1- related protein kinase (SnRK1) was identified as a potential interacting protein of TaTCTP. The results of yeast two-hybrid assay showed that TCTP could interact with SnRK1 in yeast, and the yeast carrying TCTP and SnRK1 could grow on SD/-Leu/-Trp/-His/-Ade (SD/-LWHA) medium. The fluorescence signal of the interaction between TCTP and SnRK1 was found to be distributed in the cytoplasm in the Bi-fluorescense complementation experiment. Co-IP experiments further showed that TCTP and SnRK1 could interact in plant cells. This study lays an important foundation for further studying the mechanism of TaTCTP in the interaction between wheat and Puccinia triticina, and it play a great influence on further improving the molecular mechanism of wheat resistant to Puccinia triticina.
Basidiomycota ; Humans ; Neoplasms ; Protein Biosynthesis ; Protein-Serine-Threonine Kinases ; Triticum

Databases, Protein ; Information Dissemination ; Internet ; Protein Engineering ; methods ; Protein Structure, Tertiary ; Recombinant Proteins ; biosynthesis ; chemistry

OBJECTIVETo study relation of the expression of hTERT mRNA and cyclin A, p53 protein, proliferation cell nuclear antigen (PCNA) in ameloblastoma (AB) and to investigate clinical biological characteristics of AB.

METHODSThe hTERT mRNA was detected by in situ hybridization, cyclin A, p53 protein and PCNA by SP method. Normal oral mucosa and odontogenic keratocyst (OKC) are comparition.

RESULTSThe positive ratio of hTERT mRNA was 1/7 cases in normal oral mucosa. The expression of cyclin A, p53 and PCNA in normal oral mucosa were similar, and the positive cells distributed in stratum basale. In OKC, the positive cells distributed in stratum basale and super-strrtum basale. And the positive ratio of hTERT, cyclin A, p53, PCNA in OKC was 14/16 cases, 4/32 cases, 11/25 cases, 5/9 cases, respectively. In AB, the positive ratio of hTERT mRNA, cyclin A, p53 protein and PCNA was 94.4% (51/54), 66.7% (40/60), 85.7% (42/49) and 78.1% (25/32), respectively. A strong correlation was found between hTERT mRNA with cyclin A, p53 protein and PCNA (r(s) = 0.914, 0.848, 0.804, P < 0.05).

CONCLUSIONSThe expression of hTETR mRNA, cyclin A, p53 and PCNA is different in different tissues and lesions, and correlates with cell differentiation and clinical biology behaviour. Telomerase activity is related to cumulation of p53 protein, related to cell proliferation and differentiation, regulated by cyclin A, and higher in S phase.

Ameloblastoma ; metabolism ; pathology ; Cyclin A ; biosynthesis ; Humans ; Jaw Neoplasms ; metabolism ; pathology ; Proliferating Cell Nuclear Antigen ; biosynthesis ; RNA, Messenger ; biosynthesis ; Telomerase ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis

Yarrowia lipolytica lipase Lip2 (YlLip2) is an important industrial enzyme with many potential applications. To alleviate the dissolved oxygen (DO) limitation and improve YlLip2 production during high-cell density fermentation, the YlLip2 gene lip2 and Vitreoscilla hemoglobin (VHb) gene vgb were co-expressed in Pichiapastoris under the control of AOX1 and PsADH2 promoter, respectively. The PsADH2 promoter from Pichia stipitis could be activated under oxygen limitation. The SDS-PAGE and CO-difference spectrum analysis indicated that VHb and YlLip2 had successfully co-expressed in recombinant strains. Compared with the control cells (VHb-, GS115/9Klip2), the expression levels of YlLip2 in VHb-expressing cells (VHb+, GS115/9Klip2-pZPVT) under oxygen limitation were improved 25% in shake-flask culture and 83% in a 10 L fermentor. Moreover, the VHb+ cells displayed higher biomass than VHb- cells at lower DO levels in a 10 L fermentor. In this study, we also achieved a VHb-expressing clone harboring multicopy lip2 gene (GS115/9Klip2-pZPVTlip2 49#), which showed the maximum lipolytic activity of 33 900 U/mL in a 10 L fermentor under lower DO conditions. Therefore, it can be seen that expression of VHb with PsADH2 promoter in P. pastoris combined with increasing copies of lip2 gene is an effective strategy to improve YlLip2 production.
Bacterial Proteins ; biosynthesis ; genetics ; Fermentation ; Fungal Proteins ; biosynthesis ; genetics ; Lipase ; biosynthesis ; genetics ; Oxygen ; analysis ; pharmacology ; Pichia ; metabolism ; Protein Engineering ; Recombinant Proteins ; biosynthesis ; genetics ; Truncated Hemoglobins ; biosynthesis ; genetics

Cell-free protein synthesis (CFPS) systems based on crude cell extracts have been used in protein expression in vitro. With the researchers' endeavor for decades, the CFPS system has been developed as an important research tool in many frontiers of fundamental and applied biology because of its clear genetic background and simplicity to control the reaction. The yield of CFPS systems derived from prokaryote or eukaryote has increased to several grams per liter with constantly decreasing cost. Nowadays grams of protein could be prepared using a large-scale cell-free system. Recently, the advantages on the expression of complicated, toxic and membrane proteins have shown the great potential of the CFPS systems. The rapid progress of this technology made us to believe that it will take an important place in biopharmaceutical industries undoubtedly.
Bioengineering ; trends ; Cell-Free System ; Drug Industry ; trends ; Protein Biosynthesis

TIR (Translation Initiation Region) efficiency is very important in prokaryotic expression. The TIR's efficiency is highly dependent on SD (Shine-Dalgarno) sequence, distance between SD sequence and start codon, DB (Downstream Box) sequence, TIR's second structure, codon adaptation and so on. In this paper, we designed and implemented the software to optimize DB sequence and 5' rare codons. It generated some optimization sequences by analyzing the target sequence and comparing it with 16S RNA. And the optimization sequences is sorting by number of base pairing, location of base pairing and codon adaptation. We drew up the algorithm and the core of code in this paper.
Base Sequence ; Codon ; genetics ; Programming Languages ; Protein Biosynthesis ; genetics ; Software

Eukaryotic translation initiation factor 4G (eIF4G) is a scaffold component of eukaryotic translation initiation factor 4F (eIF4F) complex, which takes principal part in the initiating of protein synthesis. Both two subtypes (eIF4G1 and eIF4G2) of eIF4G were found to be closely related with various tumors. The eIF4G1 expression is significantly up-regulated in breast cancer, cervical cancer, nasopharyngeal carcinoma, lung squamous cell carcinoma, prostatic carcinoma and other malignant tumors, compared with those in adjacent tissues; and the eIF4G2 is obviously over-expressed in diffuse large B cell lymphoma and acute myeloid leukemia, but low-expressed in bladder transitional cell carcinoma. This paper reviews the progress in the study of the role of eIF4G in tumor genesis, development, diagnosis and prognosis.
Eukaryotic Initiation Factor-4G ; Humans ; Neoplasms ; Protein Biosynthesis ; Up-Regulation

OBJECTIVETo investigate the expression of androgen receptor (AR) isoforms in normal human prostate.

METHODSFourteen normal prostatic specimens from donors, aged 25 on average (21-28 yr), were analyzed by high resolution isoelectric focusing (IEF). The expression of AR isoforms was demonstrated in all 14 normal human prostatic tissues.

RESULTSFour types of AR isoforms were detected with isoelectric point value at 6.5, 6.0, 5.8 and 5.3 in 14 prostatic specimens. Binding of 3H-dihydrotestosterone (DHT) to these four AR isoforms was inhibited by the addition of 100-fold excess of DHT and testosterone. No effect of progesterone, estradiol and diethylstilbestrol on tritiated hormone binding was observed.

CONCLUSIONSThere are four AR isoforms in normal human prostate. The expression of AR isoforms is different from one another.

Adult ; Humans ; Isoelectric Focusing ; Isoelectric Point ; Male ; Prostate ; metabolism ; Protein Isoforms ; biosynthesis ; Receptors, Androgen ; biosynthesis