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Chinese Journal of Agricultural Biotechnology最新文献

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Isolation and culture of bone marrow mesenchymal stem cells from human fetus and their biological properties 人胎骨髓间充质干细胞的分离培养及其生物学特性研究
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002404
Zhang Yihua, Dou Zhong-ying, Shen Wen-zheng, Yang Chun-rong, Gao Zhimin
{"title":"Isolation and culture of bone marrow mesenchymal stem cells from human fetus and their biological properties","authors":"Zhang Yihua, Dou Zhong-ying, Shen Wen-zheng, Yang Chun-rong, Gao Zhimin","doi":"10.1017/S1479236208002404","DOIUrl":"https://doi.org/10.1017/S1479236208002404","url":null,"abstract":"The population doubling number(70～80 times) of human fetal bone marrow mesenchymal stem cells(BMMSCs) is about two times more than that (30～40 times) of the adult BMMSCs, and their differentiation capacity is superior to that of their adult counterparts. In this study, BMMSCs were isolated from 2- to 3-month-old human abortuses by scissoring their long bones lengthwise, followed by rinsing and culturing whole marrow cells. Basic medium and serum concentration for BMMSCs culture were optimized and growth curves made, both with MTT(3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide) reduction assay. Isolated cells were identified with flow-cytometry and immunocytochemistry for their antigen markers. The biosafety of isolated cells was evaluated by karyotype analysis and tumor forming experiment. The results indicated that lengthwise scissoring of fetal long bones and rinsing of their marrow cells was practical and useful to obtain the BMMSCs from human abortuses at the age of 2～3 months. In this experiment, α-MEM(minimum essential medium alpha medium) +20% FCS(fetal cattle serum) was the best system for the BMMSCs in vitro. The third passage BMMSCs expressed Oct4, SSEA3 and SSEA4 beside the surface markers of their adult counterparts. The population doubling time of the BMMSCs of passage 6, 12 and 24 were 34,36 and 40 h, respectively. The cells in all the passages showed diploid karyotype and formed no tumor in the nude mice. The BMMSCs of human abortuses at the age of 2～3 months were proved to be biologically safe and ideal seed cells for researches on human tissue engineering and regeneration medicine.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"182 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123005723","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: 6
Cloning, expression and bioactivity of chicken receptor activator of NF-κB ligand ( chRANKL ) 鸡NF-κB配体受体激活因子(chRANKL)的克隆、表达及生物活性研究
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002131
Wang Yan, Hou Jia-fa
{"title":"Cloning, expression and bioactivity of chicken receptor activator of NF-κB ligand ( chRANKL )","authors":"Wang Yan, Hou Jia-fa","doi":"10.1017/S1479236208002131","DOIUrl":"https://doi.org/10.1017/S1479236208002131","url":null,"abstract":"Using reverse transcription polymerase chain reaction (RT-PCR) and gene splicing by overlap extension (SOE-PCR), the DNA sequence encoding the chicken's receptor activator of NF -κB ligand (RANKL) was amplified, and then cloned into the expressing plasmid pET32a(+) with His-tag, and was highly expressed in Echerichia coli. Then the purified protein was added in primary cultures of chicken osteopclasts to observe its bioactivity. The results showed that the size of PCR product was 1 200 bp which was consistent with the expected one, and the relative molecular weight of induced protein was 64 kD. Western blot indicated that the induced protein could react with anti-His antibody. It was also found that the induced protein could stimulate mature chicken osteoclasts to resorb bone.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"42 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133688728","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: 1
Activation tagging library construction and mutant phenotype analysis 激活标记文库构建及突变体表型分析
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002325
Cao Dongmei, Fan Xi-Ying, Wang Yunshan, Kang Lifang
{"title":"Activation tagging library construction and mutant phenotype analysis","authors":"Cao Dongmei, Fan Xi-Ying, Wang Yunshan, Kang Lifang","doi":"10.1017/S1479236208002325","DOIUrl":"https://doi.org/10.1017/S1479236208002325","url":null,"abstract":"Activation tagging plays an important role in plant genomics studies. In this paper, an activation tagging library containing 50 000 Basta-resistant lines was constructed by mediated transformation of the mutant bzr1-D of Aribidopsis thaliana . Among these transformants, 47 lines showed obvious phenotypes, including late flowering time, dwarf growth habit, changed leaf shape, longer leaf petiole, leaves lacking trichomes, sterility and no kink between stem/leaf. Some T-DNA flanking sequences were obtained by TAIL (thermal asymmetric interlaced) and nested PCR. Results showed that the mutants contained one to three T-DNA insertions. The insertions were distributed in the first, fourth and fifth chromosomes of the genome.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"13 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124547163","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: 1
Cryopreservation of boar semen using straws 用稻草冷冻保存野猪精液
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002301
Gao Jun-feng, Zhen Xiao-feng, Ge Li-jun, L. Qing, R. Rong
{"title":"Cryopreservation of boar semen using straws","authors":"Gao Jun-feng, Zhen Xiao-feng, Ge Li-jun, L. Qing, R. Rong","doi":"10.1017/S1479236208002301","DOIUrl":"https://doi.org/10.1017/S1479236208002301","url":null,"abstract":"An optimal protocol for cryopreservation of boar semen was established. First, the boar semen was pre-diluted with ZORLESCO (ZO) solution and pre-equilibrated at room temperature for 1 h. After adding extender I, spermatozoa were equilibrated at 5°C for 1.5 h; then an equal volume of extender II was added and the spermatozoa equilibrated for 2 h. The resulting spermatozoa were loaded into 0.25 ml straws, equilibrated for 10 min at 3 cm above the surface of liquid nitrogen (LN), then promptly submerged into LN. When thawing, straws were incubated in a water bath at 37°C for 30 s. This procedure yielded the highest post-thaw motility of 0.58±0.03 and plasma integrity of 63.2±1.2%, together with a normal acrosome in 51.4±2.6% of spermatozoa. Abnormal spermatozoa after freezing represented only 14.0±3.0%.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"33 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124307572","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: 4
SSR markers for identification of purity of melon hybrids 甜瓜杂种纯度鉴定的SSR标记
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S147923620800243X
Li Ju-fen, M. Guo-bin, Xu Ling
{"title":"SSR markers for identification of purity of melon hybrids","authors":"Li Ju-fen, M. Guo-bin, Xu Ling","doi":"10.1017/S147923620800243X","DOIUrl":"https://doi.org/10.1017/S147923620800243X","url":null,"abstract":"The hybrid purity of melon ( Cucumis melo L.) was tested by polymerase chain reaction (PCR) assay based on simple sequence repeat (SSR) markers in two F 1 melon hybrids (‘Dongfangmi 1’ and ‘Dongfangmi 2’) and their parental lines. Twelve pairs of SSR primers for ‘Dongfangmi 1’ and three pairs for ‘Dongfangmi 2’ were selected. Results showed that self-inbred seeds were effectively distinguished from F 1 hybrid seeds using these SSR primers, a finding that was consistent with the results recorded from field tests. ‘Dongfangmi 1’ and ‘Dongfangmi 2’ were identified from their parental lines, and seven other uterine hybrid lines by multiplex primers MS48+MS60 and MS4+MS20, respectively. Contamination of F 1 hybrid seeds caused by self-inbred and other unknown pollens can be effectively and more reliably detected by PCR assays with multiplex SSR primers.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130067325","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: 7
Bovine interferon-tau expression in Escherichia coli and identification of its biological activities 牛干扰素-tau在大肠杆菌中的表达及其生物活性鉴定
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S147923620800226X
Gao Fang-fang, Wu Zhongyi, Zeng Shen-ming
{"title":"Bovine interferon-tau expression in Escherichia coli and identification of its biological activities","authors":"Gao Fang-fang, Wu Zhongyi, Zeng Shen-ming","doi":"10.1017/S147923620800226X","DOIUrl":"https://doi.org/10.1017/S147923620800226X","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"22 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129171225","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: 3
Identification of the cry gene in Bacillus thuringiensis strain WZ-9 and its toxicity against Henosepilachna vigintioctomaculata
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002453
Song Ping, Wang Qin-ying, Wu Hui-xian, Lu Xiu-jun, W. Yong
{"title":"Identification of the cry gene in Bacillus thuringiensis strain WZ-9 and its toxicity against Henosepilachna vigintioctomaculata","authors":"Song Ping, Wang Qin-ying, Wu Hui-xian, Lu Xiu-jun, W. Yong","doi":"10.1017/S1479236208002453","DOIUrl":"https://doi.org/10.1017/S1479236208002453","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"123 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124155026","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: 9
Induction of lamb follicular development and embryo production in vitro 羔羊卵泡发育的体外诱导及胚胎产生
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002428
C. Xiaoyong, Tian Shujun, Sang Run-zi, Sun Shu-chun, Zhao Zhu-jun, L. Junjie, Dun Weitao
{"title":"Induction of lamb follicular development and embryo production in vitro","authors":"C. Xiaoyong, Tian Shujun, Sang Run-zi, Sun Shu-chun, Zhao Zhu-jun, L. Junjie, Dun Weitao","doi":"10.1017/S1479236208002428","DOIUrl":"https://doi.org/10.1017/S1479236208002428","url":null,"abstract":"Effect of lamb age, transport stimulation and repeated hormone superovulation on the number of collected oocytes were determined, and effect of the culture medium containing ethylene-diamine-tetra-acetic acid (EDTA) on the development of produced embryos from lamb oocytes in vitro were studied. Results indicated that the mean number of collected and available oocytes from per 6- to 8- week lamb were 60.8±13.9 and 58.2±12.3 respectively, which were more than that those (27.3±5.1 and 26.0±4.9 ) from per 12- to 14- week lamb(P 0.05), respectively; The transport stimulation didn't decreased the number of collected oocytes from the superovulation lambs(P 0.05), however, the number of collected oocytes in the repeated superovulation group was reduced significantly compared with the control group(P 0.05); The embryonic culture medium containing 10 μmol/L EDTA could improve significantly the development capability of in vitro produced embryo from lamb oocytes (P 0.05), and the healthy lambs were born by the embryo transfer.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"221 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131515225","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: 3
Comparison of Conventional Plasmid Vector and Semliki forest virus-derived Vectors in Expressing Growth Hormone Releasing Hormone (GHRH) 传统质粒载体与塞姆利基森林病毒衍生载体表达生长激素释放激素的比较
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002313
Ren Xiao-hui, Lu Hu-ying, Liu Song-cai, Z. Mingjun, O. Song-ying, Li Hong-Yi, Zhang Yong-liang
{"title":"Comparison of Conventional Plasmid Vector and Semliki forest virus-derived Vectors in Expressing Growth Hormone Releasing Hormone (GHRH)","authors":"Ren Xiao-hui, Lu Hu-ying, Liu Song-cai, Z. Mingjun, O. Song-ying, Li Hong-Yi, Zhang Yong-liang","doi":"10.1017/S1479236208002313","DOIUrl":"https://doi.org/10.1017/S1479236208002313","url":null,"abstract":"The elements for Semliki forest virus (SFV) RNA replicon, a kind of new generation vector, comes from the Alphavirus genus. It was designed to overcome the poor efficacy of some current plasmid vector. Genes coding for viral replicases are reserved while genes coding for structure proteins are replaced by foreign gene in RNA replicon. High level replication of RNA and expression of foreign gene in cytoplasm are regulated by the replicases. To evaluate the effects of the SFV RNA replicon on the efficiency of gene expression, LacZ gene was inserted into pIRES-neo which digested by BamHⅠand dephosphorylated by shrimp alkaline phosphatase, and pIRES-neo-LacZ vector was constructed. RNA replicon vector pCMV-rep-LacZ and two conventional CMV promoter-based vector (pLNCX-LacZ and pIRES-neo-LacZ) were transfected by Lipofectin to prepared 293 cells, respectively. RNA replicon vector pCMV-Rep-GHRH(Growth hormone releasing hormone) and two conventional CMV promoter-based vector(pCDNA3.1(+)-GHRH and pIRES-neo-GHRH) were transfected by Lipofectin to prepared 293 cells, respectively, too. Results of quantitating for β-galactosidase expressed from transfected cells and quantitating for GHRH expressed from transfected cells by RIA and RT-PCR showed that the express level of RNA replicon vector was superior to ordinary vector and 2~3 times higher than normal plasmid vector. This will provide a beneficial exploring to improve the efficiency of gene expression in eukaryotic cell.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"104 ","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"120939593","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: 1
In vitro differentiation of chicken spermatogonial stem cells into adipocytes 鸡精原干细胞向脂肪细胞的体外分化
Chinese Journal of Agricultural Biotechnology Pub Date : 2008-12-01 DOI: 10.1017/S1479236208002416
Yu Fei, Geng Jian-hui, Ni Li-gang, He Xian-hong, Xu Qi, Li Bichun
{"title":"In vitro differentiation of chicken spermatogonial stem cells into adipocytes","authors":"Yu Fei, Geng Jian-hui, Ni Li-gang, He Xian-hong, Xu Qi, Li Bichun","doi":"10.1017/S1479236208002416","DOIUrl":"https://doi.org/10.1017/S1479236208002416","url":null,"abstract":"","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131719169","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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