| With the development of agri-biotechnology, the industrialization of genetically modified(GM) crops is more and more extensive, transgenic breeding has become a main approach of crop breeding. In the genetically modified crops, the genetic stability of transgenic traits and expression level stability of foreign genes have been gotten special attention of breeding experts and are the key factors of security applications. In this paper, Southern blot and ELISA were used as main test methods to analyze the integration situations of new glyphosate-resistant genes and insect-resistant gene in transgenic cotton genome. The genetic stability and expression level of Bt in different generations were also studied.The main results of this research were listed as follows:1.Southern blot was used to study the genetic stability of insect-resistant gene in transgenic cotton varieties of 2000-2. The southern blot results of genome which digested by Hind â…¢ and EcoR â… separately showed that all insect-resistant plants have a single insertion site of foreign insect-resistant expression cassette, and the expression cassette can be inherited to the progeny. This integration situation ensured the stability of heredity.2.The expression level of insect-resistant protein in varieties of SCRC series and 2000-2 was tested by using ELISA. Overall, the insect-resistant protein can express in high level in SCRC and 2000-2, although it exhibit difference in different varieties. This results indicated that the insect-resistant protein can be expressed stably.3.Southern blot was used to detect the insertion situations of foreign gene in synchronous expression of the GAT and GR79. In the total 9 independent transgenic lines, 4 lines have double insertion sites and 5 lines have a single insertion site.4.A series of methods to do southern blot of cotton has been summarized based on using different probes to detect foreign gene. |
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