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cloning 2

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Turbot (Scophthalmus maximus L.) Neuropeptide Y Starvation Feeding Differential expression 1

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Study on Family Collection, Gene Mapping, Gene Identification and Gene Function of Human Genetic Diseases

Xia Jiahui

Strategic Study of CAE 2000, Volume 2, Issue 11,   Pages 1-11

Abstract: neurological deafness gene, was identified using a novel strategy of “Gene Family-Candidate Disease Gene Cloning

Keywords： genetic disease     gene mapping and cloning     gene familiy-candidate disease gene cloning     genome wide scan    

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Factors affecting early embryonic development in cattle: relevance for bovine cloning

Yanna DANG, Kun ZHANG

Frontiers of Agricultural Science and Engineering 2019, Volume 6, Issue 1,   Pages 33-41 doi: 10.15302/J-FASE-2018228

Abstract: Incomplete epigenetic reprogramming is believed to be a major limitation to cloning efficiency.., Trichostatin A) could greatly improve cloning efficiency in both mice and cattle.

Keywords： bovine cloning     embryo development     somatic cell nuclear transfer     X-inactive specific transcript    

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Full length cDNA cloning and expression of luteinizing hormone (LH) and which serum concentration was

Liu Xuezhou,Shi Bao,Wang Shanshan,Xu Yongjiang,Li Xiaoxiao

Strategic Study of CAE 2014, Volume 16, Issue 9,   Pages 50-60

Abstract:

The full length cDNA of LH has been cloned from pituitary of half smooth tongue sole C. Günther by using RACE technology for the first time. The cDNA of LH is 670 bp，and the open reading frame is 477 bp encoding peptides of 158 amino acid. Multiple amino acid sequences alignments were performed among vertebrates，and the results showed that tongue sole LH mature peptide was the most similar to their orthologs of Pleuronectiformes and Perciformes which was 58 %～68 % identity. And 12 conserved cysteine residues and 1 N-linked glycosylation site were found in the deduced amino acid sequence. Real-time PCR analysis showed that LH mRNA was expressed in all twelve tissues including brain，pituitary，ovary and so on. The highest expression level was in pituitary. And the stronger amplification signals were detected in the brain and gonad than all other non- pituitary tissues. It suggested that LH may had the more physiological functions because LH mRNA of tongue sole expressed widely in non-pituitary tissues. Quantitative real- time PCR was used for analyzing the expression of female C. semilaevis LH mRNA in the reproductive cycle. The results showed that LH mRNA was expressed in all three tissues including brain，pituitary and ovary from the stage Ⅱto stage Ⅵ， and the highest expression level appeared on the stage Ⅳ and stage Ⅴwhich suggested that the main function of LH is to promote eggs finally mature and discharge. Radio immunoassay （RIA）marked by 125I was used to detect the concentration of plasma LH in C.semilaevis. The results showed that the biggest plasma LH concentration appareed on the stage of Ⅴwhich also suggested that the main function of LH was to promote eggs finally mature and discharge.

Keywords： Cynoglossus semilaevis Günther     LH     cDNA cloning     Quantitative real-time PCR     RIA    

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Intracellular Strain Evaluation-Based Oocyte Enucleation and Its Application in Robotic Cloning Article

Mingzhu Sun, Yaowei Liu, Maosheng Cui, Qili Zhao, Xiangfei Zhao, Yidi Zhang, Jingjing Huang, Guizhang Lu, Xin Zhao

Engineering 2023, Volume 24, Issue 5,   Pages 73-83 doi: 10.1016/j.eng.2022.04.016

Abstract:

Since the first cloned sheep was produced in 1996, cloning has attracted considerable attention becauseThis low success rate is considered to be the major limitation in the extensive application of cloningWe then developed a robotic batch SCNT system to apply this micromanipulation method to animal cloningThe success rate of cloning was significantly increased compared to that of traditional methods (2.5%In addition to the cloning technique, the intracellular strain evaluation-based enucleation method is

Keywords： Oocyte enucleation     Robotic cloning     Intracellular strain     Intracellular damage     Optical flow    

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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

Frontiers of Agricultural Science and Engineering 2019, Volume 6, Issue 1,   Pages 61-65 doi: 10.15302/J-FASE-2018227

Abstract: crucial procedure for mammalian somatic cell nuclear transfer (SCNT), especially for domestic animal cloninghigh enucleation rate but has minimal negative effects on the reprogramming potential of oocyte for cloningIn this study, we compared the pig cloning efficiency resulting from blind aspiration-based (BA-B) enucleation

Keywords： cloning     enucleation     pig     SCNT    

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Embryo-mediated genome editing for accelerated genetic improvement of livestock

Zachariah MCLEAN, Björn OBACK, Götz LAIBLE

Frontiers of Agricultural Science and Engineering 2020, Volume 7, Issue 2,   Pages 148-160 doi: 10.15302/J-FASE-2019305

Abstract: It integrates embryo-based genomic selection with precise multi-editing and uses embryonic cloning with

Keywords： animal breeding     cattle     cloning     CRISPR/Cas9     cytoplasmic injection     embryo     genome editing     germline chimaeras    

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Cloning of human XAF1 gene promoter and assay of its transcription activity in a variety of cell lines

Qiong CHEN, Qing YU, Yuhu SONG, Peiyuan Li, Ying CHANG, Zhijun WANG, Lifeng LIU, Wei WU, Jusheng LIN

Frontiers of Medicine 2009, Volume 3, Issue 2,   Pages 148-152 doi: 10.1007/s11684-009-0032-7

Abstract: To investigate the regulation of tumor suppressor XAF1 gene expression in digestive system cancers, we studied XAF1 gene promoter transcription activity and mRNA level in digestive system cancer cell lines (human hepatoma cell line HepG2, human colon cancer cell line LoVo, and human gastric cancer cell line AGS) and nontumor cell lines (human embryonic liver cell line L02 (L02 cells) and human embryonic kidney 293 cells [HEK293 cells]) as controls. 1395-bp-promoter fragment of XAF1 gene was amplified by polymerase chain reaction (PCR) and cloned into pGL3-basic vector and pEGFP-1 vector to assay its promoter transcription activity. The plasmids were transfected into a variety of cell lines by lipofectamine 2000. The promoter transcription activity was determined by dual-luciferase report assay, and enhanced green fluorescent protein (EGFP)-positive cells were detected by fluorescence microscope. The expression of XAF1 mRNA in HEK293 and L02 were significantly higher than that in any of the three digestive system cancer cell lines. The dual-luciferase reporter assay showed that the promoter transcription activity in digestive system tumor cell lines transfected with pGL3-XAF1p promoter was apparently lower than that of both HEK293 and L02 cells. Expression of green fluorescent protein (GFP) under the control of XAF1 promoter in the three digestive system cancer cell lines was lower than that of both HEK293 and L02 cells. The activities of pGL3-XAF1p in the three digestive system cancer cell lines after treatment with heat stress were significantly lower than those in the unstressed cells. The results suggested that remarkably down-regulated XAF1 mRNA expression in digestive system cancer cell lines may be due to loss of transcription activity of XAF1 promoter.

Keywords： gene     X-linked inhibitor of apoptosis protein associated factor-1 (XAF1)     promoter     transcription regulation    

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cDNA Cloning of Neuropeptide Y gene of turbot Scophthalmus maximus L. and its expression analysis of

Han Jian,Liu Bin,Jiang Zhiqiang,Mu Xiaosheng,Lei Jilin

Strategic Study of CAE 2014, Volume 16, Issue 9,   Pages 42-49

Abstract:

In this study, the full length of turbot NPY cDNA was cloned using RT-PCR and rapid-amplification of cDNA ends (RACE). Turbot NPY cDNA is 729bp with a 5´-untranslated region (UTR), 135bp 3´-UTR and 299bp ORF. The open reading frame encodes a 99 amino acid NPY. The first 28 amino acids constitute the signal sequence followed by the 36 amino acid mature peptide. Turbot NPY share a 53.9–98.0% identity with NPY from other fish, especially, Turbot NPY share a 97–98.0% identity with NPY from other flounder (Japanese flounder, winter flounder).Using real-time PCR to analysis NPY expression in turbot. Tissue distribution studies show that NPY mRNA is expressed in brain, in particular the hypothalamic, and in peripheral tissues. During food deprivation, the NPY undergo significant change in expression (P<0.05), with NPY levels being elevated first, then decreased, and being significantly increased, maintain a high level of expression, then decreased again during food deprivation. Expression level was significantly higher with refeeding after fasting than the normal feeding, the longer of food deprivation, the higher of expression level. But the expression level of 2h after refeeding will fall again to a similar level. Our results suggest that NPY is conserved peptides that might be involved in the regulation of feeding and other physiological functions in turbot.

Keywords： Turbot (Scophthalmus maximus L.) Neuropeptide Y Starvation Feeding Differential expression    

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Factors influencing the somatic cell nuclear transfer efficiency in pigs

Yong JIN, Manling ZHANG, Xinrong JU, Shuang LIANG, Qiang XIONG, Lihua ZHAO, Xiaowei NIE, Daorong HOU, Qiang LIU, Junzheng WANG, Chenyu WANG, Xiaokang LI, Lining ZHANG, Xiaorui LIU, Ying WANG, Haiyuan YANG, Yifan DAI, Rongfeng LI

Frontiers of Agricultural Science and Engineering 2019, Volume 6, Issue 1,   Pages 73-80 doi: 10.15302/J-FASE-2018220

Abstract:

Using a data set from our laboratory, we assessed the effects of several factors on pig cloning efficiencyconfluence (>90%) used as donor cell resulted in higher pregnancy rate, delivery rate and overall cloningof blastocysts transferred by somatic cell nuclear transfer (SCNT) did not significantly affect the cloning

Keywords： blastocyst     donor cell     estrus synchronization     pregnancy rate     pig cloning     somatic cell nuclear transfer    

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Title Author Date Type Operation

Study on Family Collection, Gene Mapping, Gene Identification and Gene Function of Human Genetic Diseases

Xia Jiahui

Journal Article

Factors affecting early embryonic development in cattle: relevance for bovine cloning

Yanna DANG, Kun ZHANG

Journal Article

Full length cDNA cloning and expression of luteinizing hormone (LH) and which serum concentration was

Liu Xuezhou,Shi Bao,Wang Shanshan,Xu Yongjiang,Li Xiaoxiao

Journal Article

Intracellular Strain Evaluation-Based Oocyte Enucleation and Its Application in Robotic Cloning

Mingzhu Sun, Yaowei Liu, Maosheng Cui, Qili Zhao, Xiangfei Zhao, Yidi Zhang, Jingjing Huang, Guizhang Lu, Xin Zhao

Journal Article

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

Journal Article

Embryo-mediated genome editing for accelerated genetic improvement of livestock

Zachariah MCLEAN, Björn OBACK, Götz LAIBLE

Journal Article

Cloning of human XAF1 gene promoter and assay of its transcription activity in a variety of cell lines

Qiong CHEN, Qing YU, Yuhu SONG, Peiyuan Li, Ying CHANG, Zhijun WANG, Lifeng LIU, Wei WU, Jusheng LIN

Journal Article

cDNA Cloning of Neuropeptide Y gene of turbot Scophthalmus maximus L. and its expression analysis of

Han Jian,Liu Bin,Jiang Zhiqiang,Mu Xiaosheng,Lei Jilin

Journal Article

Factors influencing the somatic cell nuclear transfer efficiency in pigs

Yong JIN, Manling ZHANG, Xinrong JU, Shuang LIANG, Qiang XIONG, Lihua ZHAO, Xiaowei NIE, Daorong HOU, Qiang LIU, Junzheng WANG, Chenyu WANG, Xiaokang LI, Lining ZHANG, Xiaorui LIU, Ying WANG, Haiyuan YANG, Yifan DAI, Rongfeng LI

Journal Article