地高辛标记LEA3蛋白基因探针的制备及其在转基因草莓核酸杂交中的应用Preparation of LEA3 Protein Gene Probe Labeled by Digoxigenin and Application in Detection of Transgenic Strawberry
王俊丽,葛会波,彭士琪,张红梅,续九如
摘要(Abstract)：
用地高辛对胚胎发生晚期丰富蛋白LEA3基因片段(1.0 kh)进行了探针标记。斑点杂交表明,该探针的检测灵敏度为5 Pg。用地高辛标记和未标记的标准分子量DL 2000进行杂交时,只检测到2.00、1.00、0.75、0.50 kb4条杂交带。应用LEA3探针对转基因草莓(Fragaria ananassa Duch.)进行了Southern blot分析,检测到1.0 kh杂交带,表明外源LEA3基因整合到草莓染色体上。
关键词(KeyWords)： 地高辛;LEA3基因;探针标记;转基因草莓
基金项目(Foundation): 河北省自然科学基金资助项目(302110)。
作者(Author): 王俊丽,葛会波,彭士琪,张红梅,续九如
DOI: 10.13925/j.cnki.gsxb.2004.04.012
参考文献(References)：
- [1] Jimenez-Bermudez S, Redondo-Nevado J, Munoz-Blanco J, et el. Manipulation of strawberry fruit softening by antisense expression of a peetate lyase gene[J]. Plant Physiol, 2002, 128: 751-759 .
- [2] Chen G, Ye CM, Huang JC , et al. Cloning of the papaya ringspot virus (PRSV) replicase gene and generation of PRSV-resistant papayas through the introduction of the PRSV repliease gene[J]. Plant Cell Reports, 2001, 20(3): 272-277.
- [3] Zhu L, Holefors A, Ahlman A, et al. Transformation of the apple rootstock M.9/29 with the rolB gene and its influence on rooting and growth[J]. Plant Sci, 2001, 160(3): 433-439.
- [4] Gao M, Tao R, Miura K, et al. Transformation of Japanese persimmon (Diospyros kaki Thunb.) with apple cDNA encoding NADP-dependent sorbitol-6-phosphate dehydrogenase[J]. Plant Sci, 2001, 160(5): 837-845.
- [5]郭北海,张艳敏,李洪杰,等.甜菜碱醛脱氢酶(BADH)基因转化 小麦及其表达[J].植物学报,2000,42(3):279~283.
- [6] Guo KJ, Scott Bowden D. Digoxigenin-labeled probe for the detection of hepatitis B virus DNA in serum[J]. Journal of Clinical Microbiology, 1991,29(3): 506-509.
- [7] Xu DP, Duan XL, Wang BY, et al. Expression of a late embryogen esis abundant protein gene, HVA1, from barley confers tolerance to water deficit and salt stress in transgenic rice[J]. Plant Physiol, 1996, 110:249-257.
- [8]华志华,黄大年.转基因植物中外源基因的遗传学行为[J].植物 学报,1999,41(1):1-5.