In plant genetic engineering, selectable marker is needed to distinguish transformant. As the commercialization of transgenic plants, people are more and more paying close attention to their safety, among which mainly refers to the safety of selectable marker. In order to increase the safety of transgenic plants, biologists began to search for biosafe selectable marker.Current research progress of taking key enzyme in metabolic pathways were overviewed,which include glycometabolism, amino acid metabolism, hormone metabolism ,nucleotide metabolism and protein metabolism et cetera, as selectable marker in transgenic plants.

OBJECTIVE: To suppress NgR gene expression in neural stem cells and observe differentiation of neural stem cells in vitro after interfered which provide nutritional support for the facial nerve repair in vivo. METHOD: PCR amplification, restriction endonuclease digestion, T4DNA ligase connections were used to connected NgR with rector pGCsi, and constructed recombinant vector (NgR shRNA). Lipofectamine 2000 were used to transfect the NSC. The expression of NgR was examined by Western Blot. The proportion of neural stem cells transformed into neurons after transfection was tested by Immunocytochemistry. Neural stem cells were planted in PLGA tubes after transfected, and were scanned by electron microscopy. RESULT: NgR shRNA plasmid was constructed and infected neural stem cells successfully. Western Blot showed that the expression of NgR decreased in neural stem cells after interference. Immunocytochemistry showed that the rate of the neural stem cells transformed into neurons after interfered was significantly higher (P < 0.01). CONCLUSION Neural stem cells were transformed into neurons after NgR shRNA plasmid infected neural stem cells, which promoted axonal regeneration more effectively and provided a efficient and stable gene platform for facial nerve repair.
Animals ; Cell Differentiation ; Cells, Cultured ; Facial Nerve ; surgery ; GPI-Linked Proteins ; genetics ; metabolism ; Myelin Proteins ; genetics ; metabolism ; Neural Stem Cells ; cytology ; metabolism ; Nogo Receptor 1 ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism

The rodipicolinate synthase gene was cloned from Nicotiana tabacum L.cv,which encodes the key enzyme in the synthesis of lysine,and then a vitro molecular reform reliefing feedback suppression was made. Some resistant seedlings were acquired using the mutated gene as selectable marker and analog of lysine as selectable reagent,and the detection results were positive by means of PCR and Real-Time PCR,however the phenotype is somewhat abnormal.

Female ; Head and Neck Neoplasms ; pathology ; Humans ; Infant ; Teratoma ; pathology

Aged ; Apoptosis ; physiology ; Carcinoma, Squamous Cell ; genetics ; Cell Cycle ; Female ; Gene Expression Profiling ; Humans ; Laryngeal Neoplasms ; genetics ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Signal Transduction

Objective To optimize and establish the methodology for culturing and inducing differentiation of mouse preadipocytes 3T3-L1. Methods The mouse cells 3T3-L1 were incubated in DMEM medium contained with 10%FBS, during which the incubation medium was refreshed every 2 to 3 days. Two methods were used to introduce differentiation, including three-drug combination group and four-drug combination group. The protocol of mediumⅠin three-drug combination group including insulin 10 mg/L, IBMX 0.5 mmol/L and DEX 1.0μmol/L. The protocol of mediumⅠin four-drug combination group including indometacin 0.1 mmol/L based on those of three-drug combination group. Both of them were incubated for 2 days and continuous for 2 times. And medium Ⅱ included insulin 10 mg/L for 2-day culturing and continuous for 2 times. Oil red O staining was used to observe the morphological changes of two groups of cells before and after treatment under inverted microscope. Results Mouse preadipocytes 3T3-L1 appeared in good conditions and grew in a paving stone fashion. These cells covered homogeneously the bottom of incubators, the culture medium refreshed every 2 days. The results of four-drug combination group were better than those of three-drug combination group. After three-drug combination induced differentiation, there was no significant change in cell morphology. Comparing with three-drug combination induced differentiation, four-drug combination was successfully achieved in over 90% of the cell inducing, which were round-shaped, with jacinth ester droplets by oil-red O staining. Conclusion We have optimized the method for culturing and inducing differentiation of mouse preadipocytes 3T3-L1 by adding indometacin on the basis of the three-drug combination induced differentiation.

Pseudomonas sp. TS1138 isolated from soil samples was able to form L-cysteine from DL-2-Amino-△2-Thia-zoline-4-Carboxylic Acid (DL-ATC) after cultured 16 hours . The optimum carbon and nitrogen soruces of strain growth and enzyme formation are glucose and urea. This enzyme was induced by DL-ATC. The product was identified to be L-Cysteine based on thin layer chromatography, optical rotation and HPLC studies.

Objective To explore the clinical value of renal dynamic imaging and urinary N-acetyl-?-D-glucosaminidase(NAG),apoptosis DNA fragment(ADF) in evaluating the damage degree of hydronephrotic kidneys(HnK) in children with hydronephrosis.Methods Level of glomerular filtration rate(GFR) was detected in 41 children with congenital hydronephrosis by renal dynamic imaging,and urine NAG,ADF in pelvis in HnK and healthy kidneys (as controls) were detected by enzyme-linked immuno-sorbent assay(ELISA).Patholo-gic changes of HnK in 41 children were graded intoⅠ~Ⅴ according to Elder standard. And GFR,urinary NAG and ADF of HnK were divi-ded into subgroup according to pathologic changes ,at the same time statistical analysis was performed within each groups. And the correlations of pathologic grades with GFR,urinary NAG and ADF of HnK were analyzed.Results 1.Kindneys GFR in healthy kidneys and Hnk were (174.33?20.43)?10-3 L/min,(143.86?17.51)?10-3 L/min respectinely,and there was significant difference between healthy kidneys and Hnk (P0.05).3.There was significant negative correlation between GFR levels of HnK and pathologic grades(r=-0.814 P0.05).Conclusions For hydronephrotic kidneys,urinary NAG can eva-luate impaired nephric tubule whereas renal dynamic imaging may evaluate the damage level of glomeruli;urine ADF may not indicate the damage level of diseased kidneys in children with congenital hydronephrosis.

Objective To study the resistance and molecular profiles of Staphylococcus aureus strains isolated from the clinical specimens.Methods Antimicrobial susceptibility was tested with 56 strains of Staphylococcus aureus isolated from a hospital from May to November 2011.The mecA and pvl genes were detected.The spa genetic types were analyzed.Results A total of 21 (37.5%)Staphylococcus aureus strains were resistant to methicillin (MRSA)and 35 (62.5%)were sensitive to methicillin (MSSA).Nineteen of the 21 (90.5%)MRSA strains carried mecA gene.Compared with MSSA,MRSA were much less sen-sitive to rifampin,fluoroqunolones,tetracycline and gentamicin (P <0.05).All the MRSA isolates were susceptible to vanco-mycin,linezolid,tigecycline,quinupristin-dalfopristin and nitrofurantoin.Six spa types were identified among the MRSA strains.Type t030 was the most prevalent,accounting for 66.7% (14/21)of all the MRSA strains.MRSA-t030 and MRSA-t002 were resistant to multiple antibiotics.Eighteen spa types were identified among the MSSA strains.Type t189,t377 and t034 were the top three spa types of MSSA,accounting for 14.3%,14.3% and 11.4%,respectively.A new MSSA spa typ-ing strain new1 was isolated from pus.There were five Panton-Valentine leukocidin (PVL)-positive isolates,3 of which was MSSA-t189 type. Conclusions Type t030 is the most prevalent spa type among clinical MRSA strains,which is resistant to many kinds of antibiotics and widely spreads in the hospital setting.There are many different spa types a-mong the MSSA strains.Type t389,t377 and t034 are the top three spa types of MSSA.