Mining of candidate genes involved in the biosynthesis of dextrorotatory borneol in Cinnamomum burmannii by transcriptomic analysis on three chemotypes

Background Dextrorotatory borneol (D-borneol), a cyclic monoterpene, is widely used in traditional Chinese medicine as an efficient topical analgesic drug. Fresh leaves of Cinnamomum trees, e.g., C. burmannii and C. camphor, are the main sources from which D-borneol is extracted by steam distillation, yet with low yields. Insufficient supply of D-borneol has hampered its clinical use and production of patent remedies for a long time. Biological synthesis of D-borneol offers an additional approach; however, mechanisms of D-borneol biosynthesis remain mostly unresolved. Hence, it is important and necessary to elucidate the biosynthetic pathway of D-borneol. Results Comparative analysis on the gene expression patterns of different D-borneol production C. burmannii samples facilitates elucidation on the underlying biosynthetic pathway of D-borneol. Herein, we collected three different chemotypes of C. burmannii, which harbor different contents of D-borneol.A total of 100,218 unigenes with an N50 of 1,128 bp were assembled de novo using Trinity from a total of 21.21 Gb clean bases. We used BLASTx analysis against several public databases to annotate 45,485 unigenes (45.38%) to at least one database, among which 82 unigenes were assigned to terpenoid biosynthesis pathways by KEGG annotation. In addition, we defined 8,860 unigenes as differentially expressed genes (DEGs), among which 13 DEGs were associated with terpenoid biosynthesis pathways. One 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and two monoterpene synthase, designated as CbDXS9, CbTPS2 and CbTPS3, were up-regulated in the high-borneol group compared to the low-borneol and borneol-free groups, and might be vital to biosynthesis of D-borneol in C. burmannii. In addition, we identified one WRKY, two BHLH, one AP2/ERF and three MYB candidate genes, which exhibited the same expression patterns as CbTPS2 and CbTPS3, suggesting that these transcription factors might potentially regulate D-borneol biosynthesis. Finally, quantitative real-time PCR was conducted to detect the actual expression level of those candidate genes related to the D-borneol biosynthesis pathway, and the result showed that the expression patterns of the candidate genes related to D-borneol biosynthesis were basically consistent with those revealed by transcriptome analysis. Conclusions We used transcriptome sequencing to analyze three different chemotypes of C. burmannii, identifying three candidate structural genes (one DXS, two monoterpene synthases) and seven potential transcription factor candidates (one WRKY, two BHLH, one AP2/ERF and three MYB) involved in D-borneol biosynthesis. These results provide new insight into our understanding of the production and accumulation of D-borneol in C. burmannii.

A Rational System of Design Patent Remedies

A design patent owner who wins her suit is entitled to the defendant'sentire profit from the sale of the product, whether or not the design wasthe basis for buying the product. No other IP regime has this rule, and itmakes no sense in the modern world, where a design may cover only a smallcomponent of a valuable product. The culprit is section 289 of the PatentAct, a provision added in the nineteenth century, when design patents werevery different than they are today. We should abolish section 289 and bringrationality to design patent remedies.

Means and Meaning in Patent Remedies

Professor Sichelman's article on "Purging Patent Law of Private Law Remedies" offers a welcome and useful perspective on the reform of patent remedies. In this comment I critique some of his assumptions regarding the "private" nature of patent and other remedies, then turn to discussing several examples of existing and underutilized tools, such as "reverse liability" rules, that might accomplish much of Professor Sichelman's agenda without necessarily fomenting a radical re-conceptualization of patent remedies doctrine. Along the way I suggest how deployment of such tools might play out in a variety of current patent controversies, including Sichelman's example of patent "trolls," as well as FRAND licensing and pharmaceutical "pay for delay" agreements.

Patent Remedies

Since the Supreme Court's eBay decision in 2006, the U.S. has employed a hybrid patent remedy system that mixes property and liability rules. When the patent owner and the infringer are competitors, the courts typically issue a permanent injunction requiring the infringer to cease its infringing activities. In contrast, when the patent owner and the infringer are not competitors, the courts often allow the infringer to continue its infringing activities so long as it pays specified ongoing royalties to the patent holder. This article studies the incentives for invention and for new product development in such a hybrid system.