幽門螺旋桿菌(以下簡稱HP)是一種革蘭氏陰性螺旋狀桿菌，坐落在胃中，全球估計有一半的人受過感染，由於缺乏抗菌的療法，HP可以長期存活在宿主體內，使得罹患下列疾病的機率大增，例如，12指腸潰瘍或胃潰瘍，胃腺癌，或粘膜相關淋巴組織淋巴瘤；在過去的研究中，發現HP可以加強胃上皮細胞激增，或減弱細胞的自殺(apoptosis)能力，這些情形都有可能會增加罹患胃癌或相關疾病的風險，在過去幾年間，解釋HP基因體的資訊有相當大的進展，基因體的研究可以了解HP的基因與發病的關係，而目前HP 基因體 26695 strain 已經被完整定序出來，它是由胃炎的病人上所取出的菌體，雖然現在已有許多在Hp基因體上的研究，但卻很少在蛋白質體或是結構基因體上的研究。
目前在結構基因體計畫中已完成了以下的工作:我們從HP中選擇了20個基因來做結構的研究，這些基因分別為: HP1144, HP1215, HP1423, HP0892, HP0496, HP0495, HP0274, HP0902, HP0199, HP0817, HP0222, HP0385, HP1049, HP1219, HP1324, HP1425, HP1492, HP0032, HP0458, 和 HP0187。由所選出的20個基因中，進行聚合酵素連鎖反應，經過多次調整黏合溫度及反應時間，分別在黏合溫度 57oC做出HP1144，HP1215，HP1423，HP0892，HP0496，HP0495，HP1219，HP1425 ，HP1324，HP1049，HP0385，HP0222，53oC則做出HP0274，HP0902，HP0817，HP0032，HP0458，HP1492，HP0187，HP0199，然將這些標的物轉植至pET22b載體，HP0495, HP0222, HP0385, HP0187, HP1492, HP0458, HP1324已成功的表現出蛋白質，HP0385, HP0495,也已純化完成。

旋光儀在結構基因體中可以提供二級結構的分析，而從旋光儀光譜，可以了解蛋白質的穩定性，HP0385, HP0495,在熱變性實驗中，熔點分別為70.08oC,85.72oC，顯示出其具有相當高的熱穩定性，在化學變性實驗，HP0495化學變性的中點為6.18M，表現出相當高的蛋白質穩定性。而後由濃縮時所產生的現象和1H-15N HSQC光譜推斷HP0385 在高濃度時會產生聚集的現象，最後選擇HP0495並且取得15N和13C標定的蛋白質在600MHz核磁共振儀進HNCA, HNCO, HN(CO)CA, HN(CA)CO, CBCANH, CBCA(CO)NH三組三維的實驗，並利用三組光譜來進行主鏈的序列分析。

Helicobacter pylori is a Gram-negative bacterium that colonizes the stomachs of an estimated half of all humans. In the absence of antimicrobial therapy, H. pylori maintains residency in the stomach for the remainder of its host’s lifespan. In a subset of humans, H. pylori infection leads to serious disease such as duodenal or gastric ulcer, gastric adenocarcinoma, or mucosa-associated lymphoid tissue lymphoma. It was also found that H. pylori can enhance gastric epithelial cell proliferation and attenuate apoptosis in vivo, which may partially explain the increased risk of gastric cancer associated with this disease. During the past years, a great deal of progress has been made in defining genetic variation among H. pylori isolates. The tools and approaches used to address this problem illustrate how the current revolution in genomics is affecting the study of microbial pathogenesis. The genomes 26695 strain were sequenced in their entirety, 26695 having been isolated from a patient with gastritis. Although lots of efforts have been made on the studies of genomics of H. pylori, little has been done on the proteomics especially structural genomics studies of H. pylori.
We have accomplished the following work for this project: Twenty of the H. pylori genes were chosen for structural studies.These genes are HP-1144, HP-1215, HP-1423, HP-0892, HP-0496, HP-0495, HP-0274, HP-0902, HP-0199, HP-0817, HP-0222, HP-0385, HP-1049, HP-1219, HP-1324, HP-1425, HP-1492, HP-0032, HP-0458, and HP-0187. The PCR products could be amplified in the presence of 1-10 ng genomic DNA template at annealing temperatures of: 57 oC - HP-1144, HP-1215, HP-1423, HP-0892, HP-0496, HP-0495, HP-0222, HP-0385, HP-1049, HP-1219, HP-1324, and HP-1425; 53 oC - HP-0274, HP-0902, HP-0199, HP-0817, HP-1492, HP-0032, HP-0458, and HP-0187. We are subcloning these targets into pET22b and HP-0495, HP-0222, HP-0385, HP-0187, HP-1492, HP-0458, HP-1324 was expressed successfully .Now Hp-0385, HP-0495, HP-1492, HP-0458, were purified.CD may provide a useful secondary screen in structure proteomics. From the CD spectra,we can better understand the contribution of protein stability to sample behavior.Thermal melting of HP-0385, HP-0495,in phosphate buffer at PH6.5 indicated unfolding of the secondary structure with Tm of 70.08oC,85.72oC, suggests that HP-0495, HP-0385, exhibits high thermostability. Urea denaturation suggests that HP-0495 exhibits high structural stability with a Cm of 6.18M. We screen HP-0385 by 1H-15N HSQC which may aggregation in high concentration. Finally, we selected HP-0495 and carried out using 15N/13C double labeled proteins on our 500 and 600-MHz NMR spectrometers. The backbone sequential assignments were determined from six triple resonance experiments (HNCA, HNCO, HN(CO)CA, HN(CA)CO, CBCANH, CBCA(CO)NH).

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