サイトメガロウイルス感染症の迅速DNA診断法の開発

The conventional diagnostic procedures for CMV infections have been both isolation of viruses and titeration of serum antibodies. These methods need several days for the final diagnosis, causing a delay in the diagnosis. Therefore, a rapid diagnostic procedure has been highly required. In this study, we conducted polymerase chain reaction (PCR) using three sets of primers corresponding IE-, LA- and V-regions of the CMV genome. Using the former two sets of primers, many bands were amplified from a DNA sample isolated from the peripheral blood leukocytes of a patient with CMV infection. However, using the latter set of primers (V-region), a single band of 305bp was amplified. Using these V-region primers, no hybridization was needed to detect the specific band. No PCR amplification products was detected by using the V-region primers from DNAs isolated from normal subjects. We also tried to use urine sedimentation samples without extraction of DNA. Sedimented cells were resuspended in water, heated by a microwave oven and used for PCR templates. A 305bp band was also detected. Subsequently, the entire procedure of our method to detect the CMV DNA requires only 4 hours. Therefore, this PCR method is reliable and useful in making rapid diagnosis of CMV infection

ヒト胎盤絨毛組織中の生長促進物質の精製

The purification of cell growth stimulation peptide from human placental chorionic tissue that controls the division of a nomal human skin fibroblast (NLF) and several human cell lines maintained in tissue culture is reported. This cell growth stimulation peptide is about 15kDa by polyacrylamide gel electrophoresis and gel chromatography. The yield of cell growth stimulation peptide is 1.2mg per 100g of human placental chorionic tissue. Growth stimulating activity of DNA synthesis in NLF cells at concentration of 0.5μg/ml is equal to activity of 10% FBS

ソバ種実から得られる低分子性タンパク質の精製と2,3の理化学的性質

Two low molecular weight proteins were separately isolated in crystalline forms from buckwheat seeds by affinity chromatography on a chitin column, ion exchange chromatography on a CM-cellulose column and gel filtration through a Sephadex G-25 column. The two crystalline proteins thus obtained were referred to as buckwheat chitin-binding proteins I and II, BCP I and II, respectively. Further purification of the crystalline BCP II by high performance liquid chromatography resulted in separation into the three proteins, BCP IIa1, IIa2 and IIb, respectively. These three purified proteins showed a single band on SDS-polyacrylamide gel electrophoresis, indicating that each protein was essentially homogeneous. The purified BCP IIa1, IIa2 and IIb had the very similar amino acid compositions, mutually, and comprised the total numbers of amino acid residues of 40 to 46, suggesting that they are closely related to each other. The amino acid compositions of the purified proteins revealed that the minimum molecular weights of BCP IIa1, IIa2 and IIb were 4,800, 5,500 and 4,800, respectively. The proteins were stable in the wide range of pH3 to 11 at lower temperature than 60℃ and even in the range of acidic pH at 100℃

ソバ種実から得られる低分子性タンパク質のキチン結合活性ならびに抗真菌活性

The low molecular weight proteins which had affinity for chitin were purified in two types of crystalline forms from buckwheat seeds and named as buckwheat chitin-binding proteins I and II, BCP I and II. respectively. To elucidate biological properties of BCP II of them, chitin-binding and antifungal activities of the protein were examined in detail. BCP II possessed the binding specificity for β-1, 4-linked N-acetylglucosamine polysaccharide, chitin, and bound to chitin, reversibly. It was found that the incubation of the protein with chitin at 30℃ and pH 8.0 for 20min was most suitable for the chitin-binding. The equilibrium absorption constant, Ka, and maximum amount of the bound protein, [PC]max, values for binding of BCP II with chitin were estimated to be 0.03 1/μmol and 18.2/μmol/g, respectively. This [PC]max value indicates that approximately 87.3mg of the protein utilizes 1g of chitin for binding. BCP II also showed the growth inhibition against fungi, Saccharomyces, Candida, Aspergillus and Penicillum tested, though the growth of bacteria was not inhibited by the protein, suggesting that the protein may play a important role as defense protein against invasion of plant pathogenic fungi into seeds

Cultural characteristics of Laetiporus sulphureus, producing an anti-thrombin substance

The clotting time for thrombin of the culture broth from Laetiporus sulphureus was more than 44 times that of the control (2% malt extract medium), whereas the culture broths of all other strains exhibited lower anti-coagulative activity. In experiments on the utilization of carbon sources, the mycelial growth of L. sulphureus was rapid with glucose as the sole carbon source. Casamino acid was the best source for the strain. Thiamine was required for the mycelial growth of L. sulphureus. The optimum temperature for efficient mycelial growth was recorded at 30℃. The initial pH 4 to 5 was the most favorable for L. sulphureus for growth