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Sun HC Xue FM Qian K Fang HX Qiu HL Zhang XY Yin ZH 《Journal of Zhejiang University. Science. B》2006,7(4):324-330
To develop a gene therapy strategy for treating bovine mastitis, a new mammary-specific vector containing human lysozyme （hLYZ） cDNA and kanamycin resistance gene was constructed for intramammary expression and clinical studies. After one time acupuncture or intracisternal infusion of healthy cows with 400μg of the p215C3LYZ vector, over 2.0 μg/ml ofrhLYZ could be detected by enzymatic assay for about 3 weeks in the milk samples. Western blotting showed that rhLYZ secreted into milk samples from the vector-injected cows had molecular weight similar to that of the natural hLYZ in human colostrums. Twenty days after the primary injection, the quarters were re-injected with the same vector by quarter acupuncture and even higher concentrations of rhL YZ could be detected. Indirect competitive ELISA of milk samples showed that the vector injection did not induce detectable humoral immune response against hL YZ. Clinical studies showed that twice acupuncture of quarters with the p215C3LYZ vector had overt therapeutic effect on clinical and subclinical mastitis previously treated with antibiotics, including disappearance of clinical symptoms and relatively high microbiological cure rates. These data provide a solid rationale for using the vector to develop gene therapy for treating bovine mastitis. 相似文献
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抑制差减杂交(Suppression Subtractive Hybridization,SSH)是一种高效鉴定和分离克隆差异表达基因的新技术。目前,该技术在分子生物学研究的各个领域得到了广泛的应用。本对SSH的技术原理、差减cDNA库构建的技术流程、常见问题分析及优缺点等作了较全面的介绍,可为研究们提供参考。 相似文献
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Construction and characterization of a cDNA library from human liver tissue with chronic hepatitis B 总被引：3，自引：0，他引：3
Chen XH Chen Z Yao HP Chen F Zhu HH Zhou HJ 《Journal of Zhejiang University. Science. B》2005,6(4):288-294
INTRODUCTION Chronic hepatitis B virus (HBV) infection is aserious clinical problem because of its wide distribu-tion and possible adverse consequences, such as he-patic decompensation, cirrhosis and/or primary livercancer (PLC). The natural course of chronic HBVinfection is characterized by a series of hepatitic flaresor exacerbations and remissions (Ganem and Prince,2004). The severity, extent, duration and frequency ofhepatic histopathological changes in hepatitic flaresare d… 相似文献
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Qin LU Jian-ping LU Xiao-dong LI Xiao-hong LIU Hang MIN Fu-cheng LIN 《Journal of Zhejiang University. Science. B》2008,9(7)
In this study the MTP1 gene, encoding a type Ⅲ integral transmembrane protein, was isolated fi'om the rice blast fungus Magnaporthe oryzae. The Mtpl protein is 520 amino acids long and is comparable to the Ytpl protein of Saccharomyces cerevisiae with 46% sequence similarity. Prediction programs and MTP1-GFP (green fluorescent protein) fusion expression results indicate that Mtpl is a protein located at several membranes in the cytoplasm. The functions of the MTP1 gene in the growth and development of the fungus were studied using an MTP1 gene knockout mutant. The MTP1 gene was primarily ex-pressed at the hyphal and conidial stages and is necessary for conidiation and conidial germination, but is not required for patho-genicity. The △mtpl mutant grew more efficiently than the wild type strain on non-fermentable carbon sources, implying that the MTP1 gene has a unique role in respiratory growth and carbon source use. 相似文献
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Identification and characterization of polyubiquitin gene from cDNA library of aspergillus fumigatus
Jata Shankar Taruna Madan Seemi Farhat Basir P. Usha Sarma 《Indian journal of clinical biochemistry : IJCB》2005,20(1):208-212
Aspergillus fumigatus (Afu) causes allergic and invasive forms of diseases in humans. In order to identify genes relevant
for pathogenesis, a total of 235 cDNA clones were randomly selected and sequenced from cDNA library of Afu. One of the partially
sequenced cDNA clones was homologous to polyubiquitin. Sequencing of the complete cDNA clone showed an open reading frame
of 912 bases. Comparison with genomic sequence of Afu using BlastN program, revealed that polyubiquitin gene comprises of
992 bases and contains one intron of 80 bases. The recombinant expression of fusion protein showed an approximately molecular
weight of 43-kDa on SDS-PAGE. The translation product of the cDNA sequence showed four tandem repeats of 76 amino acid residues
in a single polyubiquitin protein and showed 100% identity with polyubiquitin protein sequences of S. cerevisiae, N. crassa,
C. albicans, S. pombe, and M. grisae. Polyubiquitin gene is known to play important role in a variety of cellular processes
and recently have been implicated in fungal pathogenesis. Identification of polyubiquitin gene of Afu has opened up scope
to study its role in understanding Aspergillus biology and pathogenesis. 相似文献