七星瓢虫是优良的天敌昆虫,可凭滞育特性抵御逆境胁迫,提高种群生存能力。为研究七星瓢虫的滞育分子机制,构建抑制性消减杂交文库,经反向Northern斑点杂交技术进一步筛选,得到231个差异点。blastx/n比对后,正、反向文库分别得到差异表...七星瓢虫是优良的天敌昆虫,可凭滞育特性抵御逆境胁迫,提高种群生存能力。为研究七星瓢虫的滞育分子机制,构建抑制性消减杂交文库,经反向Northern斑点杂交技术进一步筛选,得到231个差异点。blastx/n比对后,正、反向文库分别得到差异表达基因64和54个,GO分类和PATHWAY分析表明,差异表达基因参与的生物过程涉及滞育的信号传导、生物合成、初级代谢和次级代谢等。文库中发现多种基因,为进一步筛选、克隆、分析滞育相关基因提供线索。着重讨论了与抗寒性相关的28 k D抗干燥应激蛋白、卵巢发育相关的Cullin 3蛋白,能量代谢相关的异柠檬酸脱氢酶的生物学功能,进而推测其与七星瓢虫滞育的相关性。展开更多
mRNA differential display technique was performed to discuss the differential expression of genes in fat tissue between introduced European and Chinese indigenous pigs. Four anchor primers in combination with five arb...mRNA differential display technique was performed to discuss the differential expression of genes in fat tissue between introduced European and Chinese indigenous pigs. Four anchor primers in combination with five arbitrary primers （20 sets in total） were used and nearly 300 bands were observed in polyacrylamide gel, among which 29 differential display bands were obtained. Twelve of 29 cDNA fragments were identified using reverse Northern dot blot, and subsequently cloned and sequenced. Eight of 12 cDNAs had no matches in GenBank and were submitted to GenBank, and the other 4 showed similarity to identified genes from GenBank. Three among 8 novel ESTs were selected to be further identified by semiquantitative RT-PCR. In our experiment, silver staining DDRT-PCR and DIG primer DNA labeling reverse Northern dot blot were used to avoid radioactive pollution. The result showed that the expressions of 5 among 8 novel ESTs were stronger in the backfat of Tongcheng pigs and the others were weaker than that in Duroc pigs. These novel ESTs were prepared for selecting genes related to adipose cells.展开更多
文摘七星瓢虫是优良的天敌昆虫,可凭滞育特性抵御逆境胁迫,提高种群生存能力。为研究七星瓢虫的滞育分子机制,构建抑制性消减杂交文库,经反向Northern斑点杂交技术进一步筛选,得到231个差异点。blastx/n比对后,正、反向文库分别得到差异表达基因64和54个,GO分类和PATHWAY分析表明,差异表达基因参与的生物过程涉及滞育的信号传导、生物合成、初级代谢和次级代谢等。文库中发现多种基因,为进一步筛选、克隆、分析滞育相关基因提供线索。着重讨论了与抗寒性相关的28 k D抗干燥应激蛋白、卵巢发育相关的Cullin 3蛋白,能量代谢相关的异柠檬酸脱氢酶的生物学功能,进而推测其与七星瓢虫滞育的相关性。
文摘mRNA differential display technique was performed to discuss the differential expression of genes in fat tissue between introduced European and Chinese indigenous pigs. Four anchor primers in combination with five arbitrary primers （20 sets in total） were used and nearly 300 bands were observed in polyacrylamide gel, among which 29 differential display bands were obtained. Twelve of 29 cDNA fragments were identified using reverse Northern dot blot, and subsequently cloned and sequenced. Eight of 12 cDNAs had no matches in GenBank and were submitted to GenBank, and the other 4 showed similarity to identified genes from GenBank. Three among 8 novel ESTs were selected to be further identified by semiquantitative RT-PCR. In our experiment, silver staining DDRT-PCR and DIG primer DNA labeling reverse Northern dot blot were used to avoid radioactive pollution. The result showed that the expressions of 5 among 8 novel ESTs were stronger in the backfat of Tongcheng pigs and the others were weaker than that in Duroc pigs. These novel ESTs were prepared for selecting genes related to adipose cells.
