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[Cloning and expression of murine alpha1,2-fucosyltransferase gene]. 小鼠α 1,2-聚焦转移酶基因的克隆与表达
Shi yan sheng wu xue bao Pub Date : 2005-12-01
Bei Lin, Makiko Saito, Masao Iwamori
{"title":"[Cloning and expression of murine alpha1,2-fucosyltransferase gene].","authors":"Bei Lin,&nbsp;Makiko Saito,&nbsp;Masao Iwamori","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>MFUT-II, a DNA fragment encoding murine alpha1,2-fucosyltransferase,was cloned by RT-PCR. The open-reading frame was ligated into mammalian expression vector pcDNA 3.1 (pcDNA3.1-MFUT-II) and was transiently transfected into COS-7 cells using a Cellphect Transfection kit. Moreover, the expression of the gene in murine tissues was analyzed with Northern hybridization showing a 3.5-Kb mRNA transcript product in several tissues. The existence of MFUT-II was detected by Southern blot. MFUT-II was found to be a new member of the murine alpha1,2-FT gene family. Its open reading frame encoded 347 amino acids with a predicted molecular mass of 39.21 kDa and exhibited sequence homology with murine H & Sec1 gene. MFUT-II also exhibited sequence homology with Human Se gene (79.0%), Rat FTB gene (89.0%) and Rabbit FT-III gene (77.0%), respectively. COS-7 cells transfected with pcDNA3.1-MFUT- II showed alpha 1,2-fucosyltransferase activity. Southern analysis revealed that MFUT-II was present in the mouse genome as a single-copy gene. In conclusion, MFUT-II was a murine Se gene.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 6","pages":"536-44"},"PeriodicalIF":0.0,"publicationDate":"2005-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25803272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[A new culture method for primordial germ cell]. 一种新的原始生殖细胞培养方法。
Shi yan sheng wu xue bao Pub Date : 2005-12-01
Xin Lu, Qing Xu, Xiao Lin Shi
{"title":"[A new culture method for primordial germ cell].","authors":"Xin Lu,&nbsp;Qing Xu,&nbsp;Xiao Lin Shi","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>To investigate the new method that the primordial germ cells (PGCs) can long time propagate, grow and keep potency of differentiation in vitro. PGCs were co-cultured with Sertoli cells (SCs) or with homologue fibroblasts from genital ridge. When co-cultured with SCs, PGCs colonies and times of transfer of culture were greater than those co-cultured with homologue fibroblasts. At present PGCs of co-cultured with SCs have succeeded in subculturing to the 51 generation. SCs can availably enhance propagating ability as well as a long period of time maintain the characteristic of PGCs.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 6","pages":"474-80"},"PeriodicalIF":0.0,"publicationDate":"2005-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25803975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Immunocytochemical localization of estrogen receptor in the oogenesis of termites]. 白蚁产卵过程中雌激素受体的免疫细胞化学定位。
Shi yan sheng wu xue bao Pub Date : 2005-12-01
Xiao Hong Su, Geng Si Xi
{"title":"[Immunocytochemical localization of estrogen receptor in the oogenesis of termites].","authors":"Xiao Hong Su,&nbsp;Geng Si Xi","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The expression of estrogen receptor(ER) was examined in the oogenesis of termites (Reticulitermes aculabialis) with immunocytochemical localization method. The results showed ER existed in oocytes at the process of differentiation and the process of growth. ER immunopositive substance was localized in the nucleus of oocytes at the differentiation stage and (the cytoplasm of oocytes at the growth stage. On the other hand,previous studies have demonstrated the procerebrum of termites could secrete FSH (Follicle Stimulating Hormone)and LH (Luteinizing Hormone) which stimulated estrogen secreting. These results provide a new morphological proof that \"brain-gonad\" participates in the regulation of the oogenesis of termites, as have been shown in mammal.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 6","pages":"545-9"},"PeriodicalIF":0.0,"publicationDate":"2005-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25803273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[The comparison study on the preparation of chromosomes of the freshwater prawn Macrobrachraium nipponense (Crustacea, Decapoda)]. [日本沼虾(甲壳纲，十足目)染色体制备的比较研究]。
Shi yan sheng wu xue bao Pub Date : 2005-10-01
Guo Ying Qian, Cai Sheng Wang, Shang Jun Yin
{"title":"[The comparison study on the preparation of chromosomes of the freshwater prawn Macrobrachraium nipponense (Crustacea, Decapoda)].","authors":"Guo Ying Qian,&nbsp;Cai Sheng Wang,&nbsp;Shang Jun Yin","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The Chromosomes of Macrobrachium nipponense had been prepared using the materials of testis or embryo with the chromosome low-osmosis method or cell low-osmosis method. After 5.5 h, the bivalents at meiotic diplotene were appeared. The comparison of the two methods for two different materials showed that the morpha of the chromosome of testis using the low-osmosis method was better than that of using the cell low-osmosis method. Contrarily, the low-osmosis was better than the cell low-osmosis for the morpha of the chromosome of embryo. The bivalents at meiotic diplotene were rod-shaped, and centromere position could be identified easily at that time. The diploid chromosomes numbers were 104. The chromosome formula was N=11M+26SM+4ST+11T. The technique of the chromosome preparation of Macrobrachium nipponense was preliminarily discussed as well.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 5","pages":"456-60"},"PeriodicalIF":0.0,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25758262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[The cloning and expression analysis of Sox9b in Cyprinus carpio]. 鲤Sox9b基因的克隆与表达分析
Shi yan sheng wu xue bao Pub Date : 2005-10-01
Qi Yan Du, Zhong Jie Chang, Feng Yu Wang, Hui Ying Hua
{"title":"[The cloning and expression analysis of Sox9b in Cyprinus carpio].","authors":"Qi Yan Du,&nbsp;Zhong Jie Chang,&nbsp;Feng Yu Wang,&nbsp;Hui Ying Hua","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Sox9 gene is one of the important transcription factors involved in developments of many tissues and organs, particularly in sex determination and chondrogenesis. In this article, the genomic DNA of Cyprinus carpio were amplified using degenerate primers. It was found for the first time that there were two versions of Sox9 in C. carpio, Sox9a and Sox9b with the same deduced amino acid sequences in the HMG box. There is one intron in this box with the length 704bp in Sox9a and 616bp in Sox9b respectively. Furthermore, a two-phase rapid amplification of cDNA ends(RACE) was used for the isolation of the full length cDNA of Sox9b. Sequence analysis revealed a 2447bp cDNA encoding a protein of 428 amino acid. HMG box of 79-amino-acid motif was confirmed to be from 96 to 174. Sequence alignment exhibited the identity rate of amino acid of Sox9b among ten kinds of animals including C. carpio, which is above 75%. The result indicated the Sox9 gene is very conservative in the progress of evolution. The tissue-specific expression of Sox9b was studied with semi-quantity RT-PCR. The result showed that this gene expresses widespread in several organs of adult C. carpio, with the level of expression in brain and testis higher than any others.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 5","pages":"397-403"},"PeriodicalIF":0.0,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25758367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[The elp3 subunit of human elongator complex ignificantly complemented the growth defects of yeast elp3delta strain]. [人拉长复合体的elp3亚基显著弥补了酵母elp3 δ菌株的生长缺陷]。
Shi yan sheng wu xue bao Pub Date : 2005-10-01
Fen Li, Qiu Ju Han, Jun Lu, Bai Qu Huang
{"title":"[The elp3 subunit of human elongator complex ignificantly complemented the growth defects of yeast elp3delta strain].","authors":"Fen Li,&nbsp;Qiu Ju Han,&nbsp;Jun Lu,&nbsp;Bai Qu Huang","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The human elongator complex was found to be very similar to the yeast Elongator both in composition and its interaction with RNA polymerase II. But little is known about its functions in vivo. In this study, we analyzed the functions of the help3, the catalytic subunit of human Elongator, using a yeast complementation system. The results indicated that help3 was able to significantly complement the growth defects of the elp3delta yeast strain under high temperature and caffeine conditions. Gene expression analysis showed that help3 significantly resumed the slow activation of the pho5 gene under the low phosphate condition, and when heat shocked it increased the expression level of ssa3 gene. The yhelp3 containing the HAT domain of human elp3 and the remainder of yeast elp3 had a higher complementation ability than human elp3, while the yhelp3HA T- with the catalytic HAT domain deleted had no complementation ability. These results implicate that human Elp3 subunit had similar functions to those of the yeast, and the HAT activity of human Elp3 was essential for its function.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 5","pages":"369-76"},"PeriodicalIF":0.0,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25758364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Transient expression of foreign ble gene in Dunaliella salina]. 外源ble基因在盐杜氏藻中的瞬时表达
Shi yan sheng wu xue bao Pub Date : 2005-10-01
Zi Xing Li, Yu Sun, Zhi Xue Liu, Ren Tao Song, Zheng Kai Xu
{"title":"[Transient expression of foreign ble gene in Dunaliella salina].","authors":"Zi Xing Li,&nbsp;Yu Sun,&nbsp;Zhi Xue Liu,&nbsp;Ren Tao Song,&nbsp;Zheng Kai Xu","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Plasmid pSP124S with ble gene was transformed into D. salina cell by electroporation. The retention and expression of foreign gene in D.salina was explored, proper parameters were established. A large amount of foreign plasmid was found to be introduced into cells. The plasmid was degraded gradually, but can be detected within 96 hours. The ble gene was efficiently transcribed from foreign promoter and transcription was maintained at least 72 hours. The ble gene can be translated correctly and so can be used as a selectable marker for the research of genetic tranformation in D. salina.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 5","pages":"411-6"},"PeriodicalIF":0.0,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25758256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[The study on the construction of expression plasmid containing rHO-1 cDNA, analysis of rHO-1 activity and function of anti-apoptosis in HUVEC]. [含rHO-1 cDNA表达质粒的构建及对HUVEC中rHO-1活性和抗凋亡功能的分析]。
Shi yan sheng wu xue bao Pub Date : 2005-10-01
Zhen Wei Xia, Qi Qi, Xue Hong Zhang, Qing Xiang Shen, Yun Zhu Li, Shan Chang Yu
{"title":"[The study on the construction of expression plasmid containing rHO-1 cDNA, analysis of rHO-1 activity and function of anti-apoptosis in HUVEC].","authors":"Zhen Wei Xia,&nbsp;Qi Qi,&nbsp;Xue Hong Zhang,&nbsp;Qing Xiang Shen,&nbsp;Yun Zhu Li,&nbsp;Shan Chang Yu","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Xenograft rejection are due to the activation of endothelial cells and the expression of a series of proinflammatory genes encoding adhesion molecules, cytokines/chemokines and procoagulant molecules,leading to endothelial cell injury and apoptosis. HO-1 which acts as a cytoprotective gene can suppress a variety of inflammatory responses to prevent graft rejection. In this study, The plasmid containing HO-1 cDNA was constructed and transfected into human vascular endothelial cells (HUVEC) for expression using DOTAP lipsomal transfection reagents. Cells were homogenized in cell culture lysis reagent (CCLR) and the activity of HO-1 was measured. The apoptosis of HUVEC induced by tumor necrosis factor (TNF)-alpha was analyzed by flow cytometry. Meanwhile, Heme and Sn-protoporphyrin (SnPP) were added respectively to evaluate the apoptosis rate of HUVEC. The results showed that the expression of HO-1 gene can be significantly up-regulated in HUVEC transfected with pcDNA3HO1. The apoptosis rate of cells treated with Heme was less than 20% but significantly increased when cells treated with SnPP, the maximum arrived at 95%. The apoptosis rate in heme induced cells was 5-20 times lower than that in SnPP inhibited cells. The apoptosis rate was a negative relation to HO-1 expression. HO-1 protein can inhibit apoptosis in HUVEC. The results provide evidence to support the essential role of HO-1 in the cytoprotective function.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 5","pages":"441-6"},"PeriodicalIF":0.0,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25758260","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Rat oocytes in early tertiary follicles can attain appropriate maturation in combination culture system. 大鼠第三次卵泡早期卵母细胞在联合培养体系中可达到适当成熟。
Shi yan sheng wu xue bao Pub Date : 2005-10-01
Gui Jun Yan, Zheng Gu, Zhao Gui Sun, Jia Ke Tso
{"title":"Rat oocytes in early tertiary follicles can attain appropriate maturation in combination culture system.","authors":"Gui Jun Yan,&nbsp;Zheng Gu,&nbsp;Zhao Gui Sun,&nbsp;Jia Ke Tso","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We have developed a \"combination culture system\", composed of primary differentiated granulosa cells, proestrus anterior pituitary lobe and delta-4-androstenedione. This cultured system effectively induce the maturation of oocytes from early tertiary follicles of diethylstilbestrol (DES)-treated rats. After DES stimulation, an average of 189 oocytes were obtained from early tertiary follicles of unilateral ovaries; 78% of these oocytes, when cultured in this culture system, extruded the first polar body, a criterion of oocyte maturation. Of the mature oocytes, the rate of normal fertilization and egg cleavage were 88% and 93%, respectively. After 96 h of culturing in vitro, 59% of the zygotes developed into morulae or blastocysts. Embryos implanted at the two-cell stage were able to develop into healthy individuals. Thus the combination culture system provides an experimental model in which the molecular mechanisms of egg maturation, fertilization, and embryonic development can be studied.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 5","pages":"432-40"},"PeriodicalIF":0.0,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25758259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
[Study on the apoptosis and its mechanism of lovo human colorectal cancer cells induced by SFPS]. [SFPS诱导lovo人结直肠癌细胞凋亡及其机制研究]。
Shi yan sheng wu xue bao Pub Date : 2005-10-01
Lin Lin Yan, Qian Liang, Ji Cheng Li
{"title":"[Study on the apoptosis and its mechanism of lovo human colorectal cancer cells induced by SFPS].","authors":"Lin Lin Yan, Qian Liang, Ji Cheng Li","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Human colorectal cancer (lovo) cells were chose to study the anti-tumor effects of SFPS and explore the significance of caspase 3 in the apoptosis of lovo cells induced by SFPS. Inhibition of the cell proliferation was measured by MTT assay. SFPS induced apoptosis of lovo cells was observed by electron microscopy, flow cytometry and DNA electrophoresis. Furthermore, the expressions of caspase 3 mRNA and pro-caspase 3 were tested by RT-PCR and Western blot. SFPS exhibited anti-proliferative activity in the dosage and time-dependent manner. After incubation for 24, 36, 48 and 72 h, the IC50 of SFPS on lovo cells was 375 mg/L, 355 mg/L, 178 mg/L and 60 mg/L, respectively. DNA ladders of lovo cells were showed on agarose gel electrophoresis and the fragments of DNA were integral of 180-200 bp 24 hours after SFPS treatment at the doses of 5 mg/L, 50 mg/L and 300 mg/L. However, mistiness DNA ladder or smear was found in lovo cells treated with 500 mg/L SFPS. When SFPS concentration was 1000 mg/l, DNA ladder disappeared and showed smear entirely. Morphological examination showed chromosomal condensation, karyotheca margination, cell shrinkage and the presence of apoptosis bodies with electron microscopy. With concentration dependent manner, the apoptosis and sub-G1 peaks were observed through flow cytometry, but the cell cycle didn't change obviously. Furthermore, the expression of pro-caspase 3 was decreased and the level of caspase 3 mRNA was increased in the time and dose-dependent manner. It suggests that SFPS may induce the apoptosis of lovo cells in vitro resulting in the inhibition of proliferation. And caspase 3 activation may participate in the processes of the apoptosis of lovo cells induced by SFPS.</p>","PeriodicalId":77395,"journal":{"name":"Shi yan sheng wu xue bao","volume":"38 5","pages":"447-55"},"PeriodicalIF":0.0,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"25758261","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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