A lactating dairy cow mammary epithelial cell(DCMEC)model is useful for studying milk synthesis and lactation in the dairy cow mammary gland.However,the cells can only maintain their original characteristics for a lim...A lactating dairy cow mammary epithelial cell(DCMEC)model is useful for studying milk synthesis and lactation in the dairy cow mammary gland.However,the cells can only maintain their original characteristics for a limited period in vitro.Thus,the proliferative capacity and lactation pattern of subcultured DCMECs need to be characterized.In the present study,subcultured DCMECs appeared to proliferate without changes in morphology or growth pattern up to the 12th passage.Subculturing had no obvious effect on the lactation capacity of the subcultured DCMEC up to the 10th passage in vitro.These results indicated that the DCMEC line could be subpassaged up to 10 times and still maintain its original milk synthesis characteristics.Moreover,DCMECs were frozen using different methods,and the method of placing the vials in the foam box was determined after cryopreservation with the best adherence of 91.2%.This method was recommended for long-term cryopreservation for DCMECs.展开更多
Now every laboratory is facing a common problem that is seriously lacking in the standard reference molecule in the GMC detecting research center. Constructing the recombinant plasmid is the urgent demand. In order to...Now every laboratory is facing a common problem that is seriously lacking in the standard reference molecule in the GMC detecting research center. Constructing the recombinant plasmid is the urgent demand. In order to overcome this problem, this study respectively cloned CrylA（B）, BAR, CP4-EPSPS, PAT and RBCL genes into PMD 18-T vector. This constructed plasmid could be used as the standard reference molecule in the qualitative detecting of the exogenous CP4-EPSPS gene, CrylA（B） gene, BAR gene and PAT gene. Identification by double restriction endonuclease digestion and identification by PCR proved that the test result was positive. All of these indicated that the established reference molecules in this study were suitable for the qualitative analysis standard of this GMC detection.展开更多
基金Supported by the National Natural Science Foundation of China(31601141)。
文摘A lactating dairy cow mammary epithelial cell(DCMEC)model is useful for studying milk synthesis and lactation in the dairy cow mammary gland.However,the cells can only maintain their original characteristics for a limited period in vitro.Thus,the proliferative capacity and lactation pattern of subcultured DCMECs need to be characterized.In the present study,subcultured DCMECs appeared to proliferate without changes in morphology or growth pattern up to the 12th passage.Subculturing had no obvious effect on the lactation capacity of the subcultured DCMEC up to the 10th passage in vitro.These results indicated that the DCMEC line could be subpassaged up to 10 times and still maintain its original milk synthesis characteristics.Moreover,DCMECs were frozen using different methods,and the method of placing the vials in the foam box was determined after cryopreservation with the best adherence of 91.2%.This method was recommended for long-term cryopreservation for DCMECs.
基金Supported by the Innovative Team Funds of Northeast Agricultural University (CXT004-3-2)Foundation of Heilongjiang Educational Committee (11511030)
文摘Now every laboratory is facing a common problem that is seriously lacking in the standard reference molecule in the GMC detecting research center. Constructing the recombinant plasmid is the urgent demand. In order to overcome this problem, this study respectively cloned CrylA（B）, BAR, CP4-EPSPS, PAT and RBCL genes into PMD 18-T vector. This constructed plasmid could be used as the standard reference molecule in the qualitative detecting of the exogenous CP4-EPSPS gene, CrylA（B） gene, BAR gene and PAT gene. Identification by double restriction endonuclease digestion and identification by PCR proved that the test result was positive. All of these indicated that the established reference molecules in this study were suitable for the qualitative analysis standard of this GMC detection.
