Ectopic expression of a maize pollen specific gene, zm401, results in aberrant anther development in tobacco

Our previous study shows that the maize zm401 cDNA is specifically expressed in maize pollens. This evidence suggests that zm401 likely functions in pollen growth and/or development. To confirm its possible involvement in pollen development, the full length cDNA of zm40l was ectopically expressed in tobacco plants under the control of a pollen specific promoter ZM13 (from maize). RT-PCR amplification demonstrated that the ZM13-driven zm401 gene was spatially expressed in tobacco pollens. It was found that all transgenic tobacco plants expressing zm401 showed various levels of sterility, ranging from abortive flower development to male sterility. Further analyses on anther development of transgenic plants indicated multiple abnormalities in the late stages of anther development. These abnormalities include lagged degradation of the tapetum and connective tissue, failed deposition of fibrous bands in endothecium cells, and aborted pollen grain development. These results strongly suggest that zm401 plays an essential role in anther development. However the exact functions of zm401 is still unclear, and further analysis of zm401 is required to determine the exact mechanism involved in anther development.     

猪链球菌2型纤连蛋白/血纤蛋白原结合蛋白 全基因的克隆及融合表达

采用发表的引物对,以猪链球菌2型（Streptococcus suis type 2）江苏分离株HA9801的基因组DNA为模板,采用PCR方法扩增纤连蛋白/血纤蛋白原结合蛋白基因fbps（GenBank登录号为AY565303）克隆于pMD-T18载体构建成载体pMD-T-fbps.经内切酶酶切和测序鉴定后,将由pMD-T-fbps内切酶切下的片段定向克隆于表达载体pET-32a（＋）,得到重组质粒pfbps.将重组质粒pfbps转化大肠杆菌（Escherichia coli）BL21株,经IPTG诱导,可高水平地表达相对分子量为83 kD的融合蛋白.配体印迹结合试验表明,表达的融合蛋白可与人纤连蛋白结合.     

SPA/SPG融合蛋白在大肠杆菌中的高效表达

葡萄球菌A蛋白（SPA）和链球菌G蛋白（SPG）的IgG结合区编码基因融合后，克隆到大肠杆菌表达载体PGEX，SPA／SPG融合蛋白（SPAG）得到高效表达。可溶性SPAG～GST产物可被谷胱甘肽亲和柱一次性纯化。用琼扩和ELISA试验测定了表达产物与不同种属动物血清反应的性能。SPAG—GST与所有SPA、SPG各自反应的动物IgG结合，对小鼠IgG的反应优于SPA或SPG。在所测试的23种动物血清中，融合蛋白与大部分哺乳动物Ig反应。     

普通烟草叶绿素含量的遗传

对普通烟草(Nicotiana tabacum L.)叶绿素含量的遗传研究表明:白肋烟与大青烟都是正常绿叶烟核基因隐性突变的产物。白肋烟与正常绿叶烟的差异主要由两对重迭基因Yb_1-yb_1和Yb_2-yb_2造成,另有一些微效基因参与作用;大青烟与正常绿叶烟的差异主要决定于一对基因D-d。因此,正常绿叶烟、大青烟和白肋烟的基因型分別为Yb_1Yb_1Yb_2Yb_2DD、Yb_1Yb_1Yb_2Yb_2dd和yb_1yb_1yb_2yb_2DD。     

多不饱和脂肪酸的营养功能及其在家禽生产中的应用

多不饱和脂肪酸是一类具有重要生物学功能的物质 ,主要包括 n-3和 n-6系列多不饱和脂肪酸 ,它们对脂类的代谢、机体的免疫、血液生化特性的影响、机体的生长发育及细胞膜功能的发挥和基因表达调控等许多方面起着重要的作用。本文主要介绍了多不饱和脂肪酸的营养功能及其作用的机理 ,并就其在家禽生产中的应用作一综述     

Alterations of cardiac sympathetic norepinephrine transporter expression in rats with volume overload

AIM: To determine the alterations of myocardial 1-adrenergic receptor (1-AR) and cardiac sympathetic norepinephrine transporter (NET) mRNA expression, which is upstream modulator of 1-AR, in rats with longterm volume overload (VOL). METHODS: Left ventricular systolic (LVSP) and end diastolic pressure (LVEDP) of rats with VOL induced by aortacaval fistula operation and control group were measured at 314,30and 60d after the operation, the mRNA at the time points was measured by RT-PCR and Northern blot analysis and quantified by densitometry. RESULTS: The cardiac sympathetic NET specific expression is in the cardiac sympathetic ganglia. Be compared with the control group, LVSP of VOL rats decreased most dramatically by 24% (P<0.05) at 3 d, after that LSVP increased gradually and were higher than control group at 60d. LVEDP increased initially compared with the control (P<0.05), the latter recovered to the control levels. RT-PCR and Northern blot showed that in VOL rats the NET mRNA did not decreased significantly from 3to 30d, and decreased remarkably at 60d after the operation (P<0.05). The pattern of 1-AR mRNA expression were similar to the NET. CONCLUSION: The results suggest decreased levels of NET mRNA may contribute to cardiac sympathetic dysfunction in heart failure due to longterm VOL, which may lead to decreased myocardial 1-AR mRNA. We conclude that the normal NET mRNA expression may play a critical role to maintain sensitivity of 1-AR to adrenergic stimuli and cardiac contractility.     

The expression of hHR21sp gene in the peripheral blood cells and bone marrow cells of patients with Fanconi anemia

AIM:To detect the expression of hHR21^spgene in normal human peripheral blood cells and in hemapoietic cell from fanconi anemia(FA) patients bone marrow after exposure to UV and gamma-irradiation.METHODS:RNA of FA hemapoietic cell and normal peripheral blood cells were harvested after UV and gamma irradiation. hHR21^sp gene expression was analyzed using RT-PCR, southern blot hybridization, phosphoimmage autoradiogram.RESULTS:The expression of hHR21^sp gene in PBL varied with the time of UV gamma-irradiation and increased obviously six hours after 80 J/M² UV-or 5 Gy gamma-irradiation. However, the expression was down-regulated in FA hemapoietic cell from FA patients indicating that hHR21^sp expression is something related to the ability of cells to repair DNA double strand break.CONCLUSION:The expression of hHR21^sp down-regulated in the FA syndrome and this might be involved in the pathogenesis of the FA.     

Effect of transfection of tumor necrosis factor α gene and its mutants on tumorigenicity of H22 tumor cells in vivo

AIM: To compare the tumorigenecity of H22 cells transfected with TNF-α gene and its mutants(secreted TNF-α mutant, S-TNF^m, transmembrane TNF-α mutant, TM-TNF^m and wild type of TNF-α, Wt-TNF) in vivo. METHODS: Three kinds of mouse liver cancer cell line H22 expressing TNF-α and its two mutants were mixed with untransfected H22 at different effector/target ratio separately. The growth of tumor was examined after injection of 2.5×10⁵(100 μL) mixed H22 tumor cells into mice. The lymphocyte infiltration in the site of tumor and the expression of Fas on tumor cells were detected by immunohistochemistry. RESULTS: The tumorigenecity of H22 cells transfected with TNF-α gene and its mutants was significantly weakened( P <0.01). Besides the cytotoxicity of the both forms of TNF, TM-TNF was found to induce the expression of death receptor Fas by tumor cells and S-TNF was shown to promote frank lymphocyte infiltration in the site of tumor. Furthermore, a transient decrease in body weight was found in mice inoculated with H22/S-TNF^m. CONCLUSION: The tumorigenecity of tumor cells was reduced by transfection with TM-TNF or S-TNF gene. It results from the cytotoxicity of TNF and their activation of tumorcidal mechanisms in vivo. TM-TNF may induce tumor cell apoptosis via the Fas pathway while S-TNF may exert its antitumor effect by recruiting and activating lymphocytes in the site of tumor.     

Molecular Cloning and Expression of IL2 cDNA from the Tibet Pig

Interleukin-2 is a vital cytokine secreted by activated T lymphocytes, and plays important role in the regulation of cellular and humoral immunity of animals. In our experiment, IL2 cDNA of the Tibet Pig was first cloned by RT-PCR from ConA-stimulated lymphocytes in the blood and subcloned into pMD-18 T vector, which then was identified with endonuclease restriction. The sequencing result showed that Tibet pig IL-2 (TPIL-2) cDNA was 503 bp long (ORF was 465 bp) (Genbank accession number: AY 294018). The recombinant prokaryotic and eukaryotic expression plasmids of the cDNA were then constructed to analyse the ability to stimulate the proliferation of porcine lymphocytes in vitro. The recombinant porcine IL-2 expressed in the prokaryotic cells was found to be of 43 kDa molecular mass, which was consistent with a 17.4 kDa protein deduced from the IL-2 cDNA sequence (glutathione S-transferase molecular mass is 26 kDa); the recombinant protein in eukaryotic cells was confirmed by use of specific rabbit anti-porcine IL-2 serum in an ELISA. The bioactivity of TPIL-2 was detected through MTT colorimetry by stimulating the proliferation of pig ConA-stimulated blasts in vitro. The results indicate that the TPIL-2 significantly promoted the proliferation of ConA-stimulated blasts of pig. This confirms that IL-2 cDNA of the Tibet pig was successfully cloned and expressed in prokaryotic and eukaryotic cells, which lays the foundation for the the preparation of specific recombinant IL-2 protein and development of novel immune adjuvants to raise the immunity of pigs against various infectious pathogens and increase the immunoprotective efficacy of vaccines.