Gene cloning and expression of cadherin in midgut of Helicoverpa armigera and its Cry1A binding regi

来源 :中国科学C辑(英文版) | 被引量 : 0次 | 上传用户:hhzj1015
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
Cadherins belong to one of the families of animal glycoproteins responsible for calcium-dependent cell-cell adhesion. Recent literatures showed that the cadherin-like in midgut of several insects served as the receptor of Bt toxin Cry1A and the variation of cadherin-like is related to insect's resistance to Cry1A. The full-length cDNA encoding cadherin-like of Helicoverpa armigera is cloned by degenerate PCR and RACE techniques and the gene was designated as BtR-harm, which is 5581 bp in full-length, encoding 1730 amino acid residues (BtR-harm was deposited in GenBank and the accession number is AF519180). Its predicted molecular weight and isoelectric point were 195.39 kDa and 4.23, respectively. The inferred amino acid sequence includes a signal sequence, 11 cadherin repeats, a membrane-proximal region, a transmembrane region and a cytoplasmic region. Sequence analysis indicated that the deduced protein sequence was most similar to the cadherin-like from Heliothis virescens with 84.2% identity and highly similar to three other lepidopteran cadherin from Bombyx mori, Manduca sexta and Pectinophora gossypiella, with the sequence identities of 60.3.6%, 57.5% and 51.0%, respectively. The cDNA encoding cadherin gene was expressed successfully in E. coli and the recombinant proteins can bind with Cry1Ac. Truncation analysis and binding experiment of BtR-harm revealed that the Cry1A binding region was a contiguous 244-amino acid sequence, which located between amino acid 1217 and 1461. Semi-quantitative RT-PCR analysis showed that BtR-harm was highly expressed in midgut of H. armigera, very low expressed in foregut and hindgut and was not expressed in other tissues. After H. armigera producing resistance to Cry1Ac, the expression quantity of BtR-harm significantly decreased in midgut of H. armigera. It is the first confirmation that BtR-harm can function as receptor of Cry1Ac in H. armigera and the binding region was located on a contiguous 244 amino acid sequence, suggesting that the decrease of expression quantity of BtR-harm is one of the main reasons for H. armigera resistance to Cry1Ac.
其他文献
The OsDREB1 gene from rice encodes a transcription factor belonging to the DREBP transcription factor subfamily. Many DREBP transcription factors regulate gene
Nongenomic effects of glucocorticoids (GC) in various cell types have been well documented, but it still remains unknown whether the mechanism also works in hip
A numerical approach to optimize dimensionless parameters of water-flooding porous media flows is proposed based on the analysis of the sensitivity factor defin
We transiently transfected pTet-on and pTRE2hyg-luciferase into the mouse embryonic stem cells (ESCs) using lipofectamine, and analyzed its inductive effect by
A soluble matrix protein P14 with an apparent molecular mass of 14.5 kDa was isolated from fragmented nacre of pearl oysters (Pinctada fucata) treated with 10%
A novel method for predicting the secondary structures of proteins from amino acid sequence has been presented. The protein secondary structure seqlets that are
Trichosanthin (TCS) is a plant toxin with ribosome-inactivating activity. TCS can be internalized by the host cells and then attacks the ribosomes resulting in
We have amplified swine myostatin (MSTN) gene by reverse transcription polymerase chain reaction (RT-PCR) and cloned it into pGEM-T Easy vector. The cloned swin
骨肿瘤中骨肉瘤(OS)和尤文肉瘤(ES)因具有较复杂的发生、发展、转移机制和不良预后而受到医学界的广泛关注。研究表明原癌基因的激活和抑癌基因的突变、凋亡信号缺失和乏氧诱导因
目的:观察脂肪因子补体C1q/肿瘤坏死因子(TNF)相关蛋白3(CTRP3)对3T3-L1脂肪细胞脂联素(APN)、瘦素(LPT)、内脏脂肪素(VFT)及爱帕琳肽(APL)等脂肪因子表达的调节效应,以及胰岛素抵抗对该调