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This study reports that wild soybean DNA was introduced into cultivar soybean through pollen tube after soybean self-fertilizing,it led to wide variation in offspring characteristics.This indicates that outside DAN segments can be directly introduced into host plant egg cells,zygotes or early embryos,some segments can insert into host cell DNA and express.The study shows this method has significant practical value for breeding. 本文利用花粉管通道在大豆自花受粉后,将野生大豆DNA导入栽培大豆,从而引起了后代性状的广泛变异。结果进一步证明了外源DNA片段直接导入受体植物卵细胞、合子或早期胚细胞,部分片段可被受体细胞DNA整合乃至表达理论的可行性和实用价值。 Bacteriophage Lambda EMBL4 was purified by density gradient centrifugation in cesium chloride Vector DNA was prepared by double digestion of purified EMBL4 DNA with Bam Hi/Sail. High molecular weight wild soybean DNA was extracted by CTAB method and was partially digested with Sau3A. The wild soybean insert DNA fragments ranging from 10 to 22 kb were recovered from agarose gel and joined with the EMBL4 vectors. The resulting recombinant DNA was packaged in vitro, and... Bacteriophage Lambda EMBL4 was purified by density gradient centrifugation in cesium chloride Vector DNA was prepared by double digestion of purified EMBL4 DNA with Bam Hi/Sail. High molecular weight wild soybean DNA was extracted by CTAB method and was partially digested with Sau3A. The wild soybean insert DNA fragments ranging from 10 to 22 kb were recovered from agarose gel and joined with the EMBL4 vectors. The resulting recombinant DNA was packaged in vitro, and 8×10 pfu were obtained. This is completely met with the theoretical value required f6r wild soybean genomic library. The library was screened by using α'-cDNA of cultivated soybean storage protein as a probe and one positive clone was obtained. 以氯化铯密度梯度离心法纯化噬菌体λEMBL4,将纯化的EMBL4 DNA用BamH1/SalI双酶切制成载体。用CTAB(十六烷基三甲基溴化铵)法提取野生大豆(种名待定)大分子DNA,Sau3A部分酶解,从琼脂糖凝胶中回收10—22kb“目的”DNA片段,与载体连接,体外包装成重组噬菌体。所得重组子值为8×10(?)pfu(噬菌斑形成单位),达到了构建野生大豆基因文库要求的理论值。以栽培大豆7S贮藏蛋白a′-cDNA作探针,用噬菌斑原位杂交法从文库中筛选出一个阳性克隆。
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