检索结果-南通市图书馆

Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo

Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2005年第2期6卷 137-141页

作者：高波孙怀昌宋成义王志跃陈芹宋红芹 School of Veterinary Medicine Yangzhou University Yangzhou 225009 China School of Veterinary Mediciney

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyle-neimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-Nl was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the β-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

关键词： Chicken ovalbumin gene regulatory regions oviduct-specific expression vector oviduct epithelium In vivo expression

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Novel cellular evidence of secretion in sperm storage microenvironment of female Chinese soft-shelled turtle

Novel cellular evidence of secretion in sperm storage microe...

引用

中国畜牧兽医学会动物解剖及组织胚胎学分会第十九次学术研讨会

作者： Muhammad Yasir Waqas Ping Yang Shakeeb Ullah Linli Zhang Qian Zhang Quanfu Li Nisar Ahmad Wei Chen Bing Chen Qiusheng Chen College of Veterinary Medicine Nanjing Agricultural University

oviduct is not merely conduit which join the uterus and ovaries,but the venue of fertilization,sperm capacitation and sperm *** storage observed in many animal species ranges from few hours to several *** are known to store sperm for period upto year or *** examined the ultrastructure of oviduct epithelium and glandular secretions in Pelodiscusis *** storage of spermatozoa was restricted to isthmus,uterus and *** of oviduct was lined with ciliated,secretory and third type of *** third type of cells was observed with abundant mitochondria,vacuoles and narrow *** the two basic cells were found which spanning the entire *** predominant were secretory cells while the others were *** prominent characteristic of ciliated cells was the presence of secretory vacuole released in the lumen in apocrine *** cells have typical biphasic granules with dark and lucent *** addition,gland cells show prominent feature including the presence of different shape,size and electron density *** synthesis of these granules are described firstly present study,Apperantly mitochondria appear to play a role in dense granule *** granules are displaced toward the apical cytoplasm when the material of the less dense portion is added more concentric layers for final maturation with the assistance of the rough endoplasmic reticulum and *** composite granules are released into the lumen by exocytosis when the less dense portion merges with the luminal *** substance leaching out of from epithelium as well as glands may contribute to keeping the sperm quiescent and alive during storage in the female reproductive tract.

关键词： Ultrastructure oviduct epithelium Secretion Stored sperm Soft-shelled turtle

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：