Ecological safety is a major consideration in the commercialization of transgenic fish. Development of sterile transgenic triploid fish through hybridization of transgenic tetraploid fish and transgenic diploid fish i...Ecological safety is a major consideration in the commercialization of transgenic fish. Development of sterile transgenic triploid fish through hybridization of transgenic tetraploid fish and transgenic diploid fish is a feasible way to solve this problem. The "all-fish" transgene, pbcAbcGHc, containing the black carp β-actin gene promoter and the open reading frame (ORF) of the black carp growth hormone (GH) gene was constructed and introduced into fertilized eggs of allotetraploid fish through microinjection. Contrast cultivation results showed that the growth rate of 150 day-old P 0 black carp GH gene transgenic allotetraploid fish was much higher than that of controls. Sixty 150 day-old transgenic allotetraploid fish were assayed by PCR for transgene integration and 90% of fish were positive for the transgene. The transgene was detected in 13 of 20 sperm samples from male transgenic allotetraploid fish. RT-PCR detected transcription of the exogenous black carp GH gene in the muscle, liver, kidney and ovaries of the largest transgenic allotetraploid fish. This study has developed P 0 black carp GH gene transgenic allotetraploid fish with a highly increased growth rate, which provides a solid foundation for the establishment of a pure line of transgenic allotetraploid fish and for the large scale production of sterile transgenic triploid fish.展开更多
The Kaposi's Sarcoma-associated Herpesvirus (KSHV)-encoded G-protein coupled receptor (vGPCR) is an oncoprotein that is implicated in KSHV-associated malignancies. We previously revealed vGPCR incorporates sulfate...The Kaposi's Sarcoma-associated Herpesvirus (KSHV)-encoded G-protein coupled receptor (vGPCR) is an oncoprotein that is implicated in KSHV-associated malignancies. We previously revealed vGPCR incorporates sulfate groups within its extracellular N-terminal tyrosine residues (Y26 and Y28) and that this tyrosine sulfation is crucial for its tumorigenicity in nude mice. hGRO-binds vGPCR in a sulfotyrosine-dependent manner and promotes its tumorigenicity through autocrine signaling. Interestingly, an unsulfated vGPCR mutant (yydd-vGPCR) attenuated the tumor growth triggered by hGRO-α . In this study, the extracellular N-terminus of vGPCR (wt-vGN) and an unsulfated vGPCR mutant (yydd-vGN) were individually secreted, expressed and purified. A radioactive labeling assay demonstrated that wt-vGN but not yydd-vGN incorporated [ 35 S]-sulfate. In nude mice, NIH3T3 cells expressing yydd-vGN but not wt-vGN could significantly inhibit the tumor growth triggered by hGRO-α . All our data support the conclusion that the unsulfated extracellular N-terminus of vGPCR reduces the tumorigenicity of hGRO-α .展开更多
We report the effective slowing and trapping of Cs atoms in an ultrahigh-vacuum apparatus.The heated Cs atoms in an oven are slowed using a Zeeman slower after the oven chamber and then trapped using a magneto-optical...We report the effective slowing and trapping of Cs atoms in an ultrahigh-vacuum apparatus.The heated Cs atoms in an oven are slowed using a Zeeman slower after the oven chamber and then trapped using a magneto-optical trap in a science chamber.Compared to the traditional vacuum pressure of~10^(−7)Pa determined by the vapor pressure of Cs atoms in the oven chamber,the designed cold nipple and differential pumping tube are used between the oven and the oven chamber to achieve a lower vacuum pressure of~3.6×10^(−9)Pa.This is beneficial for achieving and maintaining an ultrahigh vacuum in the science chamber.We demonstrate the performance of our apparatus through the effective slowing of Cs atoms and an optimal magneto-optical trap.展开更多
基金supported by the National Special Fund for Scientific Research in Public Benefits (Grant No. 20090304608)the National Basic Research Program of China (Grant No. 2007CB109006)+1 种基金the National Natural Science Foundation of China (Grant No. 30500294)the Excellent Talent Program of Hunan Normal University (Grant No. ET31004)
文摘Ecological safety is a major consideration in the commercialization of transgenic fish. Development of sterile transgenic triploid fish through hybridization of transgenic tetraploid fish and transgenic diploid fish is a feasible way to solve this problem. The "all-fish" transgene, pbcAbcGHc, containing the black carp β-actin gene promoter and the open reading frame (ORF) of the black carp growth hormone (GH) gene was constructed and introduced into fertilized eggs of allotetraploid fish through microinjection. Contrast cultivation results showed that the growth rate of 150 day-old P 0 black carp GH gene transgenic allotetraploid fish was much higher than that of controls. Sixty 150 day-old transgenic allotetraploid fish were assayed by PCR for transgene integration and 90% of fish were positive for the transgene. The transgene was detected in 13 of 20 sperm samples from male transgenic allotetraploid fish. RT-PCR detected transcription of the exogenous black carp GH gene in the muscle, liver, kidney and ovaries of the largest transgenic allotetraploid fish. This study has developed P 0 black carp GH gene transgenic allotetraploid fish with a highly increased growth rate, which provides a solid foundation for the establishment of a pure line of transgenic allotetraploid fish and for the large scale production of sterile transgenic triploid fish.
基金supported by the National Natural Science Foundation of China (81171583,31272634)the Program for New Century Excellent Talents in University (NCET-11-0971)+2 种基金the Scientific Research Foundation for the Returned Overseas Chinese Scholars/State Education Ministry(2011-1568-1)funds from Hunan Province (12JJ1005,12A088)the Excellent Talent Program of Hunan Normal University (ET31004)
文摘The Kaposi's Sarcoma-associated Herpesvirus (KSHV)-encoded G-protein coupled receptor (vGPCR) is an oncoprotein that is implicated in KSHV-associated malignancies. We previously revealed vGPCR incorporates sulfate groups within its extracellular N-terminal tyrosine residues (Y26 and Y28) and that this tyrosine sulfation is crucial for its tumorigenicity in nude mice. hGRO-binds vGPCR in a sulfotyrosine-dependent manner and promotes its tumorigenicity through autocrine signaling. Interestingly, an unsulfated vGPCR mutant (yydd-vGPCR) attenuated the tumor growth triggered by hGRO-α . In this study, the extracellular N-terminus of vGPCR (wt-vGN) and an unsulfated vGPCR mutant (yydd-vGN) were individually secreted, expressed and purified. A radioactive labeling assay demonstrated that wt-vGN but not yydd-vGN incorporated [ 35 S]-sulfate. In nude mice, NIH3T3 cells expressing yydd-vGN but not wt-vGN could significantly inhibit the tumor growth triggered by hGRO-α . All our data support the conclusion that the unsulfated extracellular N-terminus of vGPCR reduces the tumorigenicity of hGRO-α .
文摘We report the effective slowing and trapping of Cs atoms in an ultrahigh-vacuum apparatus.The heated Cs atoms in an oven are slowed using a Zeeman slower after the oven chamber and then trapped using a magneto-optical trap in a science chamber.Compared to the traditional vacuum pressure of~10^(−7)Pa determined by the vapor pressure of Cs atoms in the oven chamber,the designed cold nipple and differential pumping tube are used between the oven and the oven chamber to achieve a lower vacuum pressure of~3.6×10^(−9)Pa.This is beneficial for achieving and maintaining an ultrahigh vacuum in the science chamber.We demonstrate the performance of our apparatus through the effective slowing of Cs atoms and an optimal magneto-optical trap.
