人G6PD T279A和T279S两种突变子的体外构建

目的:构建葡萄糖6磷酸脱氢酶(Glucose 6-phosphate dehydrogenase,G6PD)T279A和T279S两种突变子.方法:以Genbank No X03674为参考序列设计并合成引物、以含G6PD基因的质粒(Philip JMason博士惠赠)为模板,PCR扩增获得G6PD野生型基因片段,琼脂糖凝胶电泳后回收PCR产物,连接、转化构建克隆质粒pMD18T-G6PD;酶切pMD18T-G6PD质粒、电泳后回收目的基因片段,连接、转化构建含G6PD野生型基因的重组质粒pAL-G6PD;设计并合成含有突变序列的引物,以pAL-G6PD为模板,体外扩增获得G6PD835-海口(835A→G,T279A)和835-中国-1(835 A→T,T279S)突变子.结果:酶切后经电泳鉴定表明获得与预期大小相符的pMD18T-G6PD质粒,EcoRI和Hind Ⅲ双酶切获得与预期大小相符的pAL-G6PD,测序结果与参考序列完全一致.0.8%的琼脂糖凝胶电泳鉴定,并定量pAL-G6PD单链DNA浓度约为200ng/uL.经测序鉴定并与参考序列比对结果表明获得了G6PD的T279A和T279S两种突变子.结论:成功构建了G6PD的T279A和T279S两种突变子,为下一步原核表达、生化性质以及酶动力学等研究奠定了基础.     

EcoRⅡ甲基化酶基因在小鼠NIH3T3细胞中的表达

CG外甲基化在哺乳动物细胞中的生物学意义仍不清楚。建立了特定位点CCWGG（W＝A／T）甲基化的哺乳动物细胞株。来自大肠杆菌的EcoRⅡ甲基化酶（催化CCWGG→CmCWGG）基因经定点诱变后重组到真核表达载体pCI－neo上,采用脂质体介导的方法将其转染小鼠NIH3T3细胞,经G418筛选获得有效表达的细胞株,酶切证实该细胞株基因组DNA上的大部分EcoRⅡ识别位点已被甲基化,PCR鉴定EcoRⅡ甲基化酶基因已稳定整合到细胞染色体上。     

汉族人群NOS3 A-922G、NOS3 T-786C 与NOS3 G894T SNP的等位基因及其组合分布与高血压的相关性

为了分析汉族人群一氧化氮合酶基因NOS3 A-922G、NOS3 T-786C 与NOS3 G894T单核苷酸多态性（single nucleotide polymorphism,SNP）的等位基因及其组合分布与高血压病的相关性,选取无亲缘关系的高血压病人192例(男97例,女95例)以及无亲缘关系的健康个体122例（男76例,女46例）为对照组,提取静脉血白细胞基因组DNA,采用等位基因特异性引物PCR技术检测NOS3 A-922G、NOS3 T-786C 与NOS3 G894T 3个位点的基因型。其结果显示：高血压病组与对照组NOS3 G894T、NOS3A-922G及NOS3 T-786C各等位基因型及其基因单倍型频率比较无显著性差异(P>0.05)。男、女性别分层研究：无论男亚组还是女亚组均未发现NOS3 A-922G、NOS3 T-786C 与NOS3 G894T各个位点SNP与高血压病有相关性。等位基因组合分布研究发现NOS3 G894G +A-922G+T-786T组合基因型总体频率分布在高血压病组与正常对照组之间有显著性差异(P<0.05,χ2= 4.5944)。男、女性别分层研究：男亚组上述3个位点SNP的各个组合基因型分布频率在高血压病组与正常对照组之间无显著性差异(P>0.05);女亚组中携带NOS3 G894G+A-922G+T-786C 的组合基因型分布频率在高血压病组与正常对照组之间有显著性差异(P<0.01,χ2=8.502)。研究发现,在中国汉族人群中NOS3A–922 G、NOS3 T-786C 与NOS3 G894T SNP与高血压病无明确的相关性,且无性别差异。组合分布研究发现,NOS3 G894G+A-922G+T-786C 的组合基因型分布频率在高血压病女性亚组较健康女性亚组明显减低,提示携带该组合基因型女性人群可能不易患高血压病。     

中华鳖MyoD1基因SNP鉴定及其与生长性状的关联分析

为了研究生肌决定因子1(MyoD1)基因多态性与中华鳖生长性状的相关性,采用直接测序法在MyoD1基因上共检测到6个SNP位点(T-49G、A-38G、C91T、A187T、C880T和T1522A),其中C880T位于外显子上,属于错义突变。对从同批繁殖、同块稻田养殖的2冬龄中华鳖群体中随机选取的178只个体中各位点的基因型进行检测。结果显示,所有位点在中华鳖群体中的平均有效等位基因数、平均观测杂合度和平均期望杂合度分别为1.636 5、0.349 3和0.375 4,除A187T位点外,其余5个位点的基因型频率分布均符合Hardy-Weinberg定律。采用一般线性模型分析各位点与中华鳖生长性状之间的相关性,研究发现,T-49G位点GG基因型个体的背甲宽显著大于TT基因型,A-38G位点AG基因型个体的背甲宽显著大于AA基因型,A187T位点TT基因型个体的体高显著大于AA基因型,T1522A位点AA基因型个体的体质量显著大于TT、TA基因型,其余位点不同基因型个体间的生长性状均不存在显著差异。T-49G、A-38G、A187T和T1522A位点与生长性状显著相关,可作为中华鳖分子标记辅助育种的候选标记。     

汉族人群NOS3 A-922G、NOS3 T-786C与NOS3 G894T SNP的等位基因及其组合分布与高血压的相关性

为了分析汉族人群一氧化氮合酶基因NOS3 A-922G、NOS3 T-786C与NOS3 G894T单核苷酸多态性(sin-gle nucleotide polymorphism,SNP)的等位基因及其组合分布与高血压病的相关性,选取无亲缘关系的高血压病人192例(男97例,女95例)以及无亲缘关系的健康个体122例(男76例,女46例)为对照组,提取静脉血白细胞基因组DNA,采用等位基因特异性引物PCR技术检测NOS3 A-922G、NOS3 T-786C与NOS3 G894T 3个位点的基因型.其结果显示:高血压病组与对照组NOS3 G894T、NOS3A-922G及NOS3 T-786C各等位基因型及其基因单倍型频率比较无显著性差异(P＞0.05).男、女性别分层研究:无论男亚组还是女亚组均未发现NOS3 A-922G、NOS3T-786C与NOS3 G894T各个位点SNP与高血压病有相关性.等位基因组合分布研究发现NOS3 G894G A-922G T-786T组合基因型总体频率分布在高血压病组与正常对照组之间有显著性差异(P＜0.05,x2=4.5944).男、女性别分层研究:男亚组上述3个位点SNP的各个组合基因型分布频率在高血压病组与正常对照组之间无显著性差异(P＞0.05);女亚组中携带NOS3 G894G A-922G T-786C的组合基因型分布频率在高血压病组与正常对照组之间有显著性差异(P＜0.01,x2=8.502).研究发现,在中国汉族人群中NOS3A-922 G、NOS3 T-786C与NOS3G894T SNP与高血压病无明确的相关性,且无性别差异.组合分布研究发现,NOS3 G894G A-922G T-786C的组合基因型分布频率在高血压病女性亚组较健康女性亚组明显减低,提示携带该组合基因型女性人群可能不易患高血压病.     

东方田鼠长江亚种和指名亚种基因组DNA序列比较分析

根据东方田鼠基因组DNA序列片段(CenBank登录号：AF277394),通过PCR方法扩增东方田鼠长江亚种(Microtus fortis calamorum)和宁夏指名亚种(Microtus fortis fortis)基因组DNA,得到-670bp左右的特异扩增片段。将PCR扩增产物克隆到pGEM-Teasy裁体,进行DNA序列分析,并用生物信息学方法比较东方田鼠长江亚种与指名亚种之间该序列的差异。结果表明：在东方田鼠两个亚种中共发现19个不同的等位基因,不同的个体在该序列存在广泛的单个核苷酸多态性(SNP),多态性位点多达25个;变换类型包括：转换(G→A、A→G、T→C、C→T)、颠换(G→T、A→T、T→A、C→A)、插入(CA)和缺失(TGTTTT)。东方田鼠长江亚种和指名亚种两个种群之间存在明显差别,尤其是在146、192、223、224、235位,但两种群间同源性仍高达98％。同时采用系统发育树(phylogenetic tree)分析方法,对两个亚种的亲缘关系进行了分析和比较,结果显示,东方田鼠长江亚种和宁夏指名亚种基因组DNA明显的分为两大组别。     

一株斑点热立克次体的病原学研究

自内蒙古自治区的草原革蜱（D．Nuttalli）中分离到一株斑点热立克次体。用微量酶标染色方法进行血清学鉴定表明,系西伯利亚（Rickettsiaesibirica种血清型。G＋Cmol％含量为32.4％。对其核酸进行聚合酶链扩增／限制性核酸内切酶消化（PCR／RFLP）分析DNA长度多态性,分别引用立氏立克次体蛋白抗原基因Rr190序列作为扩增引物Rr190．70p～602n和Rr190．4442p～5664n,扩增后分别用Pst1和Rsal限制性核酸内切酶消化。其电泳图谱证     

黑曲霉CICIM F0410中α-葡萄糖苷酶的酶学性质研究

研究了黑曲霉(Aspergillus niger)CICIMF0410发酵粗酶液中α-葡萄糖苷酶的酶学性质,发现该酶最适反应温度为55℃,最适反应pH值5.0,在65℃以下保持酶活力相对稳定,且能在pH值(4.0～6.5)范围内保持相对较高的酶活力。以该菌株的染色体DNA为模板,PCR扩增出大小为3.1kb的片段,经序列比对发现与NCBI上已公布的黑曲霉(Aspergillus niger)CBS513.88(登录号XP_001402053)序列有2个碱基的区别:2272(A→G),3098(T→G),导致2个氨基酸残基的变化:687(Asn→Ser)、983(Ser→Arg)。     

乳酸克鲁维酵母表达型T载体的研发及应用

[目的]构建以乳酸克鲁维酵母表达载体p KLAC2为基础的表达型T载体。[方法]利用定点突变技术突变载体p KLAC2上的XcmⅠ位点,获得表达载体p KL-MUT;PCR扩增含黄色荧光蛋白基因的XcmⅠ酶切盒,通过EcoRⅠ和XhoⅠ位点连接至表达载体p KL-MUT,构建成重组载体p KL-YFP,限制性内切酶XcmⅠ酶切后即产生T载体p KL-T。连接融合基因14-3-3-Zs G转化至乳酸克鲁维酵母GG799。[结果]利用该T载体可成功克隆外源基因14-3-3-Zs G并在乳酸克鲁维酵母中表达14-3-3-Zs G蛋白,荧光和Western blotting验证正确。[结论]乳酸克鲁维酵母表达型T载体已成功构建,具有快速克隆高效分泌表达外源基因的特点,对于促进乳酸克鲁维酵母表达系统表达相关蛋白的产业化具有实际意义。     

pcDNA3.1(-)-Bcr-Abl及pcDNA 3.1(-)-Bcr-Abl T3151突变质粒真核表达载体的构建

目的:将Bcr-Abl及Bcr-Abl T3151突变克隆入pcDNA3.1(-)真核表达载体,为研究靶向降解受体型酪氨酸激酶BCR-Abl,抑制肿瘤细胞生长提供研究基础.方法:pcDNA3.1 (-)-Bcr-Abl质粒构建:分别设计引物,通过分段PCR将BCR-ABL克隆入pcDNA3.1(-).首先通过PCR扩增出Bcr-a片段,将其克隆入pcDNA3.1(-)的NheⅠ/XhoⅠ之间;接着将PCR扩增出的Abl-c片段克隆入KpnⅠ/ HindⅢ之间,最后XhoⅠ/KpnⅠ双酶切pGD210,将酶切下片段插入pcDNA3.1(-)的相应位点即可.酶切鉴定及测序正确后,转染293T细胞,Western blot验证质粒的表达.pcDNA 3.1(-)-Bcr-Abl T3151的突变质粒:首先设计引物,第一步以pcDNA3.1(-)-BCR/ABL为模板,以Abl-c-u和ba-M1为引物扩增出A-1:560 bp.第二步,相同模板,以ba-M2和ba-M-down为引物扩增出A-2:870 bp.第三步,以扩增出的A-1和A-2为模板,以Abl-c-u和ba-M-down为引物,扩增出1434 bp的片段A-l+2,以Bcl和Kpn Ⅰ分别酶切pcDNA3.1 (-)-BCR/ABL以及A-1+2,将突变后的A-l+2置换入pcDNA3.1 (-)-BCR/ABL.结果:PCR结果显示3.1(-)-Bcr-Abl及3.1 (-)-Bcr-Abl T3151突变质粒条带大小符合,重组质粒经酶切鉴定和测序结果正确,转染后可见融合蛋白的表达.结论:成功构建pcDNA3.1 (-)-Bcr-Abl及pcDNA 3.1(-)-Bcr-Abl T3151的真核表达载体,并且转染293T细胞后证实其能够正确表达,为后续研究奠定了基础.     

CARD15 Gene Polymorphisms Are Associated with Tuberculosis Susceptibility in Chinese Holstein Cows

Bovine tuberculosis (BTB) is a significant veterinary and financial problem in many parts of the world. Associations between specific host genes and susceptibility to mycobacterial infections, such as tuberculosis, have been reported in several species. The objective of this study was to identify and evaluate the relationship of single-nucleotide polymorphisms (SNPs) in the CARD15 gene with susceptibility to BTB in Chinese Holstein cows. DNA samples from 201 Chinese Holstein cows (103 cases and 98 controls) were collected from Kunming City, Yuxi City, and Dali City in China. SNPs in the CARD15 gene were assessed using polymerase chain reaction (PCR) and restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). Case-control association testing and statistical analysis identified six SNPs associated with susceptibility to BTB in Chinese Holstein cows. The frequency of genotypes C/T, A/G, A/G, A/G, C/T, and A/G in E4 (-37), 208, 1644, 1648, 1799, and E10 (+107), respectively, was significantly higher in cases than in controls, and also the alleles C, A, A, G, T, and A, respectively, were associated with a greater relative risk in cases than in controls. The distribution of two haplotypes, TGGACA and CAGACA, was significantly different between cases and controls. Overall, this case-control study suggested that E4 (-37)(C/T), 208(A/G), 1644(A/G), 1648(A/G), 1799(C/T), and E10 (+107)(A/G) in the CARD15 gene were significantly associated with susceptibility to BTB in Chinese Holstein cows and that haplotypes TGGACA and CAGACA could be used as genetic markers in marker-assisted breeding programs for breeding cows with high resistance to BTB.     

Polymorphisms in genes involved in the inflammatory response and interaction with NSAID use or smoking in relation to lung cancer risk in a prospective study

Lung cancer risk was investigated in relation to single nucleotide polymorphisms in genes involved in the inflammatory response. The aim was to see if polymorphisms modifying the inflammatory response are associated with risk of lung cancer and if there were interactions between the same polymorphism and factors, which modify an inflammatory response, such as smoking status, duration, and intensity, and use of NSAID. The functional SNPs IL-1B T-31C, IL6 G-174C, IL8 T-251A, IL10 C-592T, COX2 C8473T, COX2 A-1195G and PPARgamma2 Pro(12)Ala were included. A case-cohort study including 428 lung cancer cases and a sub-cohort of 800 persons was nested within a population-based prospective study of 57,053 individuals. Variant allele carriers of IL-1B T-31C were at increased risk of lung cancer (IRR=1.51, 95% CI=1.08-2.12). There was interaction between the polymorphism COX-2 T8473C and smoking status. Thus, non-smoking variant allele carriers were at 5.75-fold (95% CI=1.25-26.43) higher risk of lung cancer than for homozygous wild type allele carriers. Lung cancer risk was similar for all genotype carriers among past and current smokers. There were, however, very few non-smoking lung cancer cases. There was interaction between IL-1B T-31C, COX-2 A-1195G and PPARgamma2 Pro(12)Ala and NSAID use in relation to lung cancer risk. For the two latter, NSAID use was only associated with a lower cancer risk among homozygous wild type allele carriers. p for interaction was 3 x 10(-6) for COX-2 A-1195G and 9 x 10(-5) for PPARgamma2 Pro(12)Ala. The results suggest that NSAID use may modify risk of lung cancer differently depending on the genotype.     

Association of +45(T/G) and +276(G/T) polymorphisms in the adiponectin gene with coronary artery disease in a population of Iranian patients with type 2 diabetes

The relation of Two single nucleotide polymorphisms (SNPs) at the adiponectin locus (+45T/G and +276G/T) with coronary artery disease (CAD) is controversial. The aim of the present study was to evaluate the genetic influence of the adiponectin gene polymorphisms in the development of CAD among patients with Type 2 diabetes (T2D). The adiponectin genotypes were detected by polymerase chain reaction and restriction analysis (PCR-RFLP) in our patients. Two adiponectin gene (ADIPOQ) SNPs (i.e. SNPs +45T>G and +276G>T) were genotyped in 114 Type 2 diabetic subjects with CAD, and 127 Type 2 diabetic patients without CAD. Demographic and anthropometric data along with plasma biochemistry including lipids, glycemic indices, and adiponectin were collected. There was a significant difference in the distribution of genotypes of +45T/G and +276G/T between CAD and non-CAD individuals (P < 0.05). Based on our results SNP+276G>T is associated with decreased risk of CAD after adjustment for potential confounding factors [adjusted OR = 0.39 (95%CI: 0.22–0.68); P = 0.001]. Similar findings were not observed for the +45T>G SNP. Two haplotypes 45T-276T and 45G-276T were associated with a decreased risk of CAD [adjusted OR = 0.47 (95% CI: 0.32–0.94); P = 0.03 and adjusted OR = 0.33 (95% CI: 0.13–0.83); P = 0.02 respectively]. No significant difference was observed between HOMA-IR, BMI, waist circumference, history of hypertension, HbA1C, and lipid concentrations regarding the two SNPs. In conclusion, these findings suggest that T allele of +276G>T SNP is significantly associated with decreased risk of CAD in T2D Patients. Also Haplotype analysis showed that two haplotypes 45T-276T and 45G-276T were associated with a decreased risk of CAD.     

SNP detection and haplotype analysis in partial sequence of MSTN gene in sheep

Polymerase chain reaction (PCR) products of the MSTN gene amplified from sixty sheep of nine Chinese indigenous sheep breeds and one imported sheep breed were sequenced to identify the single-nucleotide polymorphisms (SNPs) in a 378-bp fragment including intron 2 and exon 3 of the MSTN gene. A total of fifteen SNPs (A1937C, T1942G, C1956T, A1972C, A1990G, A2008C, A2011G, C2019T, A2025C, A2027C, T2085G, T2173C, C2198T, C2210T and C2213T) were detected among the sixty sequenced individuals and they were all located in intron 2. Twelve haplotypes were identified from these fifteen SNPs, of which haplotype I (CGTCGCGTCCGCTTT) and VIII (ATCAAAACAATTCCC) were the two major and basic ones with frequencies of 12.25% and 77.80%, respectively. Haplotype VIII was distributed in all sheep breeds and all individuals of the meat or meat-wool type sheep breeds were homozygous with respect to this haplotype. This suggests that haplotype VIII might be related to meat production traits in sheep. Haplotype I was only distributed in the fur, lambskin type and fur-meat type sheep breeds. This suggests that haplotype I may have some relationship with fur traits in sheep.     

小型猪胰淀素基因的多态性分析

目的探讨我国3个特有的小型猪品系,巴马小型猪,五指山小型猪,中国农大小型猪胰淀素（IAPP）基因多态性分布,为我国小型猪在2型糖尿病及代谢性疾病研究中的应用提供基础资料。方法提取3个品系小型猪血液基因组DNA,针对IAPP基因外显子3~内含子3的部分序列进行PCR扩增,产物鉴定、测序,统计分析基因多态。结果在所扩增的片段中共检测出2个SNPs位点,SNPs1：43G→A,位于外显子上,未引起的氨基酸的改变,突变发生在五指山猪（G/A杂合突变为16.7%,A纯合突变为83.3%）和中国农大猪（G/A杂合突变60%,A纯合突变为20%两个品系中。SNPs2：214C→T,位于内含子上,发生在五指山猪（T/C杂合突变为16.7%,T纯合突变为83.3%）和中国农大猪（T/C杂合突变为20%,T纯合突变为60%）两个品系中。结论在IAPP基因外显子3-内含子3的部分扩增序列中发现了2个SNPs位点,在3个品系小型猪中的分布不同。     

Prevalence of coagulation factor II G20210A and factor V G1691A Leiden polymorphisms in Chechans, a genetically isolated population in Jordan

Background Coagulation factor II G20210A and coagulation factor V (Leiden) G1691A single nucleotide polymorphisms (SNPs) are major inherited risk factors of venous thromboembolism. In view of the heterogeneity in their world distribution and lack of sufficient information about their distribution among Chechans, we addressed the prevalence of these SNPs in the Chechan population in Jordan, a genetically isolated population. Methods and Results factor II G20210A and factor V Leiden SNPs were analysed by polymerase chain reaction and restriction fragment length polymorphism (PCR?CRFLP) method and Amplification refractory mutation detection system (ARMS) respectively in 120 random unrelated subjects from the Chechan population in Jordan. Among the subjects studied for factor II G20210A mutation there were three individuals carrying this mutation as heterozygous (one female and two male), giving a prevalence of 2.5?% and an allele frequency of 1.25?%. No homozygous factor II allele was found. Factor V Leiden G1691A mutation was detected as heterozygous in 22 of 120 of individuals (17 female and five male) indicating a prevalence of 18.3?% and allele frequency of 9.2?%. No homozygous allele was found. Conclusion Our results indicated that prevalence of factor II G20210A mutation in the Chechan population is similar to prevalence in Jordan and Caucasian populations (1?C6?%) while the prevalence of factor V Leiden was higher in the Chechan population compared to Jordan and Caucasian populations (2?C15?%).     

First polymorphisms in JY-1 gene in cattle (Bos taurus indicus) and their association with sexual precocity and growth traits

Protein JY-1 is a bovine oocyte-specific protein that regulates granulosa cell function and is involved in early embryonic development, influencing the chance of pregnancy. This study investigated molecular markers for the JY-1 gene. Seven SNPs were identified in exon 3 of the gene. The positions of the SNPs in the exon and the respective substitutions are: 163 (T/C), 281 (T/C), 321 (T/C), 532 (T/C), 652 (A/G), 679 (T/C), and 722 (G/C) (GenBank: JN592587 and JF262042.2). SNP 163 is located in a coding region and causes a proline-to-leucine substitution. The other SNPs are located in the 3′UTR region. SNPs 163, 281, 321, and 679 were genotyped in 297 Nellore heifers and the haplotypes were constructed. The haplotypes of JY-1 were not correlated with the traits studied at 5?%.     

Association of B3GNT5 Polymorphisms with Susceptibility to ETEC F4ab/ac in the White Duroc?×?Erhualian Intercross and 15 Outbred Pig Breeds

The B3GNT5 gene is a candidate for the F4ab/ac receptor conferring susceptibility to enterotoxigenic Escherichia coli (ETEC) F4ab/ac in pigs. In this study, we screened mutations in the complete coding region of the porcine B3GNT5 gene and identified four SNPs in the 3' untranslated regions. We genotyped the four SNPs across a large-scale White Duroc × Chinese Erhualian F2 resource population (total F2 = 755) and 292 purebred piglets representing 15 Chinese and Western breeds. We found that the g.1476G→A locus and haplotypes [A;T;G;T] and [A;G;G;T] had significant association with susceptibility to ETEC F4ac in the resource population. None of the B3GNT5 polymorphisms and haplotypes was associated with susceptibility to ETEC F4ab/ac in outbred piglets. This result, together with other reports, supports the conclusion that B3GNT5 is not the responsible gene encoding the ETEC F4ab/ac receptors.     

Association of eNOS gene polymorphisms T-786C and G894T with blood pressure variability in man

The aim of this study was to evaluate the association of single nucleotide polymorphisms (SNPs) T-786C and G894T in the gene encoding eNOS with blood pressure variability (BPV) in man. Blood pressure was recorded beat-to-beat at rest three times in periods of one week (5 min, Finapres, breathing at 0.33 Hz) in 152 subjects (19-24 years). Systolic (SBPV(0.1r)/SBPV(0.1a)) and diastolic (DBPV(0.1r)/DBPV(0.1a)) blood pressure variabilities in relative (r.u.) and absolute (mm Hg(2)/Hz) units were determined by the spectral method as spectral power at the frequency of 0.1 Hz. Genotypes of both polymorphisms were detected using polymerase chain reaction and restriction analysis using enzymes Msp I and Ban II. Significant differences were observed in BPV among genotypes of T-786C SNP (p<0.05; Kruskal-Wallis), and among haplotypes of both SNPs (p<0.05; Kruskal-Wallis) as well. In T-786C SNP, carriers of less frequent allele (CC homozygotes and TC heterozygotes) showed significantly greater SBPV(0.1r) and SBPV(0.1a) compared to TT homozygotes (Mann-Whitney; p<0.05). The G894T variant showed no significant differences, but, both SNPs were in linkage disequilibrium (D'=0.37; p<0.01). Carriers of haplotype CT/CT (CC homozygotes of -786C/T and TT homozygotes of G894T) displayed significantly greater SBPV(0.1r), SBPV(0.1a) and DBPV(0.1a) compared to carriers of other haplotype combinations (Kruskal-Wallis; p=0.015, p=0.048, and p=0.026, respectively). In conclusion, the haplotype formed by less frequent alleles of both eNOS variants was associated with increased systolic and diastolic BPV in this study.