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Ming-Jie Chen

Publications and source records attributed to Ming-Jie Chen.

3 recordsLinked to original sources

A simple, rapid and effective method for total RNA extraction from Lentinula edodes.

A rapid, inexpensive and reliable method for total RNA extraction from fruiting bodies of Lentinula edodes containing large quantities of polysaccharides and secondary metabolites is described. An initial extraction step using saturated NaCl solution facilitates the separation of nucleic acids from contaminants and, after further extraction with organic solvents and precipitation with 2-propanol, total RNA of high purity and suitable for applications such as cDNA synthesis, RT-PCR and Northern blot hybridization was obtained. The procedure may also have wider applicability for total RNA extraction from the tissues of other mushrooms.

Blotting, Northern↗

Use of intersimple sequence repeats markers to develop strain-specific SCAR markers for Lentinula edodes.

Although Lentinula edodes is the second most important cultivated mushroom worldwide, most industrially cultivated strains have been identified only through traditional phenotypic analysis. Here, we report for the first time the use of sequence characterized amplified region (SCAR) markers for strain differentiation. SCAR markers were created by first generating and sequencing single intersimple sequence repeats fragments, and then designing primers based on these sequences to amplify strain-specific fragments of a certain size. One SCAR primer pair, ISL450F/R7 (amplifying a band of c. 450 bp), was designed to identify one strain of L. edodes (strain No. 7). The SCAR primer pair was then used to correctly amplify the single unique fragment from DNA samples taken from a total of 85 strains representing three separate species. Our data provide the foundation for a precise and rapid PCR-based strain-diagnostic system for L. edodes.

Base Sequence↗

[Inheritance of the foreign gene 1Ax1 in transgenic wheat (Triticum aestivum L.) with gene cassettes lacking vector backbone sequences].

Transformation with gene minimal expression cassettes without vector backbone sequence is a new transformation technique. In the current research, the inheritance of the foreign 1Ax1 gene cassettes were analyzed in transgenic plants recovered with gene cassettes lacking vector backbone sequences. The results showed that linear 1Ax1 gene cassettes were stably expressed and separated at the rate of 3 to 1 in the seeds of T1 generation. It suggested the functional copies of 1Ax1 gene cassettes were integrated in one sites, abiding by Mendelian genetic law. It revealed firstly the feasibility of the transformation technique with gene cassettes, and proved the inheritance of transgene cassettes in subsequent generations.

Gene Transfer Techniques↗