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期刊论文 3

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2019 3

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Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine

Jianwen CHEN, Kaiyuan PAN, Zhen CHEN, Biao DING, Dandan SONG, Wenbin BAO, Yunhai ZHANG

《农业科学与工程前沿(英文)》 2019年 第6卷 第1期   页码 66-72 doi: 10.15302/J-FASE-2018229

摘要:

Porcine viral diarrhea is an acute and highly contagious enteric disease of pigs that causes huge economic losses worldwide. Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are the main pathogens responsible for piglet viral diarrhea. However, currently there is no specific drug available for the effective treatment of viral diarrhea. Therefore, it is necessary to seek an effective method to diminish PEDV and TGEV infection rates. RNA interference has been applied successfully to inhibit the virus replication. It provides a potential strategy for breeding resistant pigs. In this study, four promoters and four short hairpin RNA (shRNA) vectors with LoxP sites at each end of the selectable marker genes were constructed to target PEDV and TGEV. These vectors were then transfected into porcine fetal fibroblasts, G418 resistant transfectants were confirmed by PCR and transgenic SCNT porcine blastocysts were obtained. These results have paved the way for future production of marker-free transgenic resistant to PEDV and TEGV pigs by SCNT.

关键词: piglet diarrhea     RNAi     PEDV     TGEV     transgenic resistance breeding    

Effects of enucleation method on

Chengcheng ZHAO, Junsong SHI, Rong ZHOU, Ranbiao MAI, Lvhua LUO, Xiaoyan HE, Hongmei JI, Gengyuan CAI, Dewu LIU, Enqin ZHENG, Zhenfang WU, Zicong LI

《农业科学与工程前沿(英文)》 2019年 第6卷 第1期   页码 61-65 doi: 10.15302/J-FASE-2018227

摘要:

Enucleation is a crucial procedure for mammalian somatic cell nuclear transfer (SCNT), especially for domestic animal cloning. Oocytes of domestic animals such as pigs and cattle contain dark lipid droplets that hinder localization and removal of the nucleus. Using an oocyte enucleation technique that can obtain a high enucleation rate but has minimal negative effects on the reprogramming potential of oocyte for cloning is beneficial for enhancing the outcome of SCNT. In this study, we compared the pig cloning efficiency resulting from blind aspiration-based (BA-B) enucleation and spindle imaging system-assisted (SIS-A) enucleation, and compared the pig SCNT success rate associated with BA-B enucleation and blind aspiration plus post-enucleation staining-based (BAPPS-B) enucleation. SIS-A enucleation achieved a significantly higher oocyte enucleation success rate and tended to obtain a higher full term development rate of SCNT embryos than BA-B enucleation. BAPPS-B enucleation also obtained significantly higher as well as full term development efficiency of cloned porcine embryos than BA-B enucleation. These data indicate that SIS-A and BAPPS-B enucleation are better approaches for pig SCNT than BA-B enucleation.

关键词: cloning     enucleation     pig     SCNT    

Comparison of birth weight and umbilical and placental characteristics of cloned and artificial insemination-derived piglets

Zheng AO, Chengfa ZHAO, Yanmin GAN, Xiao WU, Junsong SHI, Enqin ZHENG, Dewu LIU, Gengyuan CAI, Zhenfang WU, Zicong LI

《农业科学与工程前沿(英文)》 2019年 第6卷 第1期   页码 54-60 doi: 10.15302/J-FASE-2018249

摘要:

Somatic cell nuclear transfer (SCNT)-derived piglets have significantly higher stillbirth rate and postnatal mortality rate than artificial insemination (AI)-generated piglets. The question whether the low survival rate of SCNT piglets was related to birth weight, umbilical cord or placenta development was investigated. In this study, stillbirth rate, neonatal death rate, birth weight, umbilical cord status, placental parameters and placental gene expression patterns were compared between SCNT and AI piglets. Results showed that mortality rates at birth and during the neonatal stage of SCNT piglets were significantly higher than those of AI piglets. The incidence of abnormal umbilical cord in SCNT and SCNT-liveborn (SCNT-LB) piglets was significantly higher than in AI and AI-liveborn (AI-LB) piglets. Birth weight, placental weight, placental surface area and placental efficiency in SCNT and SCNT-LB piglets were significantly lower than those of AI and AI-LB piglets. Placental expression profiles of imprinting, angiopoiesis and nutrient transport-related genes were defective in SCNT-LB piglets compared with those in AI-LB piglets. Thus, the low survival rate of SCNT piglets may be associated with abnormal umbilical cord and placenta development. These characteristics may have resulted from aberrant expression of angiogenesis, nutrient transport, and imprinting-related genes in the placentas.

关键词: cloned     pig     death     placenta     SCNT     umbilical cord    

标题 作者 时间 类型 操作

Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine

Jianwen CHEN, Kaiyuan PAN, Zhen CHEN, Biao DING, Dandan SONG, Wenbin BAO, Yunhai ZHANG

期刊论文

Effects of enucleation method on

Chengcheng ZHAO, Junsong SHI, Rong ZHOU, Ranbiao MAI, Lvhua LUO, Xiaoyan HE, Hongmei JI, Gengyuan CAI, Dewu LIU, Enqin ZHENG, Zhenfang WU, Zicong LI

期刊论文

Comparison of birth weight and umbilical and placental characteristics of cloned and artificial insemination-derived piglets

Zheng AO, Chengfa ZHAO, Yanmin GAN, Xiao WU, Junsong SHI, Enqin ZHENG, Dewu LIU, Gengyuan CAI, Zhenfang WU, Zicong LI

期刊论文